Background Cotton is an important crop providing the most natural fibers all over the world. The cotton genomics community has utilized whole genome sequencing data to construct an elite gene pool in which functional ...Background Cotton is an important crop providing the most natural fibers all over the world. The cotton genomics community has utilized whole genome sequencing data to construct an elite gene pool in which functional genes are related to agronomic traits. However, the functional validation of these genes is hindered by time-consuming and inefficient genetic transformation methods. Thus, establishing a transient transformation system of high efficiency is necessary for cotton genomics.Results To improve the efficiency of transient transformation, we used the protoplasts isolated from the etiolated cotyledon as recipient. The enzymatic digestion buffer comprised 1.5%(w/v) cellulase, 0.75%(w/v) macerozyme, and 1% hemicellulase, osmotically buffered with 0.4 mol·L^(-1) mannitol. After 5 h of dark incubation at 25℃, uniform cotton protoplasts were successfully isolated with a yield of 4.6 × 10^(6) protoplasts per gram(fresh weight) and 95% viability. We incubated 100 μL protoplasts(2.5 × 10^(5)·m L^(-1)) with 15 μg plasmid in the solution of 0.4 mol·L^(-1) mannitol and 40% PEG 4000 for 15 min, ultimately achieving an optimal transient transfection efficiency of 71.47%.Conclusions This transient system demonstrated effective utility in cellular biology research through successful applications in subcellular localization analyses, bimolecular fluorescence complementation(Bi FC) verification, and prime editing vector validation. Through systematic optimization, we established an efficient and expedited protoplast-based transient transformation system and successfully applied this platform to cotton functional genomics studies.展开更多
Liver Transplantation (in Chinese), revised edition, edited by Professor Shu-Sen Zheng and prefaced by Professor Guo-Wei Sang, has just been published. The revised edition is mainly based on the clinical practice of t...Liver Transplantation (in Chinese), revised edition, edited by Professor Shu-Sen Zheng and prefaced by Professor Guo-Wei Sang, has just been published. The revised edition is mainly based on the clinical practice of the Liver Transplant Center, the First Affiliated Hospital, Zhejiang University and the experience展开更多
This book was written by five authors. I had met two of them, the first and third author. The first author, Steven Wayne Lingafelter, is a research entomologist with the Systematic Entomology Laboratory, USDA, based a...This book was written by five authors. I had met two of them, the first and third author. The first author, Steven Wayne Lingafelter, is a research entomologist with the Systematic Entomology Laboratory, USDA, based at the Smithsonian Institution＇s National Museum of Natural History. He has specialized on longhomed woodboring beetles for almost three decades and currently specializes on the Neotropical fauna.展开更多
Writing style is the essential issue even at the early stage the beginners who learnto read and write have to confront.From the part-Notes on reading and writing beforethe part of exercises of each lesson in English B...Writing style is the essential issue even at the early stage the beginners who learnto read and write have to confront.From the part-Notes on reading and writing beforethe part of exercises of each lesson in English Book V-VⅢ we can come to see that the ed-itors attempt to mix the content(ideas)with the corresponding techniques.This is展开更多
Genome editing is one of the most promising biotechnologies to improve crop performance.Common wheat is a staple food for mankind. In the past few decades both basic and applied research on common wheat has lagged beh...Genome editing is one of the most promising biotechnologies to improve crop performance.Common wheat is a staple food for mankind. In the past few decades both basic and applied research on common wheat has lagged behind other crop species due to its complex,polyploid genome and difficulties in genetic transformation. Recent breakthroughs in wheat transformation permit a revolution in wheat biotechnology. In this review, we summarize recent progress in wheat genetic transformation and its potential for wheat improvement. We then review recent progress in plant genome editing, which is now readily available in wheat. We also discuss measures to further increase transformation efficiency and potential applications of genome editing in wheat. We propose that, together with a high quality reference genome, the time for efficient genetic engineering and functionality studies in common wheat has arrived.展开更多
Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal ...Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR(TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35 S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.展开更多
The rapid developments of science and technology in China over recent decades, particularly in biomedical research, have brought forward serious challenges regarding ethical governance. Recently, Jian-kui HE, a Chines...The rapid developments of science and technology in China over recent decades, particularly in biomedical research, have brought forward serious challenges regarding ethical governance. Recently, Jian-kui HE, a Chinese scientist, claimed to have "created" the first gene-edited babies, designed to be naturally immune to the human immunodeficiency virus(HIV). The news immediately triggered widespread criticism, denouncement, and debate over the scientific and ethical legitimacy of HE’s genetic experiments. China’s guidelines and regulations have banned germline genome editing on human embryos for clinical use because of scientific and ethical concerns, in accordance with the international consensus. HE’s human experimentation has not only violated these Chinese regulations, but also breached other ethical and regulatory norms. These include questionable scientific value, unreasonable risk-benefit ratio, illegitimate ethics review, invalid informed consent, and regulatory misconduct. This series of ethical failings of HE and his team reveal the institutional failure of the current ethics governance system which largely depends on scientist’s self-regulation. The incident highlights the need for urgent improvement of ethics governance at all levels, the enforcement of technical and ethical guidelines, and the establishment of laws relating to such bioethical issues.展开更多
Animal models are extensively used in all aspects of biomedical research,with substantial contributions to our understanding of diseases,the development of pharmaceuticals,and the exploration of gene functions.The fie...Animal models are extensively used in all aspects of biomedical research,with substantial contributions to our understanding of diseases,the development of pharmaceuticals,and the exploration of gene functions.The field of genome modification in rabbits has progressed slowly.However,recent advancements,particularly in CRISPR/Cas9-related technologies,have catalyzed the successful development of various genome-edited rabbit models to mimic diverse diseases,including cardiovascular disorders,immunodeficiencies,agingrelated ailments,neurological diseases,and ophthalmic pathologies.These models hold great promise in advancing biomedical research due to their closer physiological and biochemical resemblance to humans compared to mice.This review aims to summarize the novel gene-editing approaches currently available for rabbits and present the applications and prospects of such models in biomedicine,underscoring their impact and future potential in translational medicine.展开更多
The recent emergence and application of engineered endonucleases have led to the development of genome editing tools capable of rapidly implementing various targeted genome editions in a wide range of species.Moreover...The recent emergence and application of engineered endonucleases have led to the development of genome editing tools capable of rapidly implementing various targeted genome editions in a wide range of species.Moreover,these novel tools have become easier to use and have resulted in a great increase of applications.Whilst gene knockout（KO） or knockin（KI） animal models are relatively easy to achieve,there is a bottleneck in the detection and analysis of these mutations.Although several methods exist to detect these targeted mutations,we developed a heteroduplex mobility assay on an automated microfluidic capillary electrophoresis system named HMA-CE in order to accelerate the genotyping process.The HMA-CE method uses a simple PCR amplification of genomic DNA（gDNA） followed by an automated capillary electrophoresis step which reveals a heteroduplexes（HD） signature for each mutation.This allows efficient discrimination of wild-type and genome-edited animals down to the single base pair level.展开更多
Cotton,an important industrial crop cultivated in more than 70 countries,plays a major role in the livelihood of millions of farmers and industrialists.Cotton is mainly grown for its fiber,an economic component that c...Cotton,an important industrial crop cultivated in more than 70 countries,plays a major role in the livelihood of millions of farmers and industrialists.Cotton is mainly grown for its fiber,an economic component that can be differentiated from its epidermal cells in the outer integument of a developing seed.Fiber length,fiber strength,and fiber fineness are three main attributes that contribute to the quality of cotton fibers.Recent advancements in genomics have identified key genes,which are the most important factors that govern these three traits,can be introduced into cultivars of interest via gene editing,marker-assisted selection,and transgenics,thus the narrow genetic background of cotton can be addressed and its fiber quality traits can be enhanced.Over the past two decades,quantitative trait loci(QTLs)have been mapped for different fiber traits,approximately 1850 QTLs have been mapped for fiber length,fiber strength,and fineness among which a few genes have been edited for quality improvement in cotton.In this background,the current review covers the development and the factors that influence these traits,along with the reported genes,QTLs,and the edited genomes for trait improvement.展开更多
The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;how...The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;however,a few mismatches are tolerated by the Cas9 endonuclease activity.An accurate and careful in silico-based off-target prediction while target selection is preferred to address the issue.These predictions are based on a comprehensive set of selectable parameters.Therefore,we investigated the possible off-target prediction and their screening in StERF3 gene-edited potato plants while developing StERF3-loss-of-function mutants using CRISPR/Cas9 approach.The 201 off-targets for the selected targets of the StERF3 gene were predicted,and 79 werefiltered as potential off-targets.Of these 79,twenty-five off-targets showed scores with defined cut-off values<0.5 and were analyzed in Sterf3-edited potato plants compared to wild-type plants.No off-targeting was found to have occurred in edited plants.展开更多
Dear Editor,Obtaining foreign DNA-free crops after genome editing is essential for agricultural applications,because of the current regulatory policies on gene-edited plants in various countries.However,it is usually ...Dear Editor,Obtaining foreign DNA-free crops after genome editing is essential for agricultural applications,because of the current regulatory policies on gene-edited plants in various countries.However,it is usually a time-consuming and labor-consuming process to select and confirm foreign DNA-free offspring,especially in woody plants with long-term life cycles and crop species with complex genomes or T-DNA multisite insertions during genome editing.展开更多
Xenotransplantation has entered the clinical phase in an effort to address the global organ shortage.However,recent clinical studies have revealed that current xenografts from gene-edited(GE)pigs still pose a risk of ...Xenotransplantation has entered the clinical phase in an effort to address the global organ shortage.However,recent clinical studies have revealed that current xenografts from gene-edited(GE)pigs still pose a risk of immune rejection and biosafety concerns.In this study,we successfully produced a large batch of 582 GE cloned(GEC)pigs with 10-(GTKO/CMAHKO/β4GalNT2KO/hCD46/hCD55/hCD59/hTBM/hCD39/hEPCR/hCD47)gene edits via gene editing and somatic cell cloning technologies,and successfully obtained the F1 generation.Phenotypic character ization of 10-GEC pigs revealed the deletion of three xenoantigens and the expression of seven human transgenes across various tissues.Digital droplet polymerase chain reaction and whole-genome sequencing revealed two cop ies of hCD46/hCD55/hCD59/hTBM/hCD39 and one copy of hEPCR/hCD47 in the pig genome with minimal off-target effects or damage to the porcine functional genes.The validation results showed that 10-GEC pigs could effectively inhibit attacks from human antibodies,complement and macrophages on porcine endothelial cells,and alleviated coagulation abnormalities between pigs and humans.Large-scale screening of pathogens revealed no evidence of 47 pathogens,including cytomegalovirus,in our 10-GEC pigs.Kidney,heart and liver xenografts from these 10-GEC pigs were transplanted into nonhuman primates(NHPs),which worked normally without hyperacute rejection(HAR).Among NHPs,the heart and liver orthotopic transplant recipients survived for 3 and 4 days,respectively,while the two kidney transplant recipients survived for 23 and 16 days,respectively.Pathological analysis showed interstitial hemorrhage and fibrosis,cellular hyperplasia with minor antibodies and complement deposition,but significantly reduced infiltration of CD68^(+)macrophages in 10-GEC pig kidney xenografts.In summary,we success fully produced specific pathogen-free 10-GEC donor pigs that resulted in effective mitigation of immune rejection upon multiorgan transplantation to NHPs.展开更多
Dear Editor,Genome editing tools are leading a revolution in plant breeding.In particular,prime editors(PEs)can install all types of base changes and small insertions/deletions at precise positions in plant genomes(An...Dear Editor,Genome editing tools are leading a revolution in plant breeding.In particular,prime editors(PEs)can install all types of base changes and small insertions/deletions at precise positions in plant genomes(Anzalone et al.,2019).PEs are by far the most powerful approach for improving traits conferred by gain-of-function point mutations.Early versions of PEs suffered from low editing efficiency,but the latest PEs can perform edits at a much higher efficiency thanks to the extensive efforts of re-searchers from around the world.Most modifications to improve PE efficiency have focused on the optimization of PE protein components and structure.展开更多
Beef and mutton production has been aided by breeding to integrate allelic diversity for myostatin(MSTN),but a lack of diversity in the MSTN germplasm has limited similar advances in pig farming.Moreover,insurmountabl...Beef and mutton production has been aided by breeding to integrate allelic diversity for myostatin(MSTN),but a lack of diversity in the MSTN germplasm has limited similar advances in pig farming.Moreover,insurmountable challenges with congenital lameness and a dearth of data about the impacts of feed conversion,reproduction,and meat quality in MSTN-edited pigs have also currently blocked progress.Here,in a largest-to-date evaluation of multiple MSTN-edited pig populations,we demonstrated a practical alternative edit-site-based solution that overcomes the major production obstacle of hindlimb weakness.We also provide long-term and multidomain datasets for multiple breeds that illustrate how MSTN-editing can sustainably increase the yields of breed-specific lean meat and the levels of desirable lipids without deleteriously affecting feed-conversion rates or litter size.Apart from establishing a new benchmark for the data scale and quality of genome-edited animal production,our study specifically illustrates how gene-editing site selection profoundly impacts the phenotypic outcomes in diverse genetic back-grounds.展开更多
Recent advances in genome editing technologies, particularly CRISPR/Cas, enable the alteration of DNA sequences to produce deletions, insertions, and substitutions in genes (Jaganathan et al., 2018), as well as large ...Recent advances in genome editing technologies, particularly CRISPR/Cas, enable the alteration of DNA sequences to produce deletions, insertions, and substitutions in genes (Jaganathan et al., 2018), as well as large or entire chromosome deletions in the genomes of plants and animals (Zhou et al., 2014;Adikusuma et al., 2017).展开更多
Regulatory T cells,a subset of CD4^(+)T cells,play a critical role in maintaining immune tolerance and tissue homeostasis due to their potent immunosuppressive properties.Recent advances in research have highlighted t...Regulatory T cells,a subset of CD4^(+)T cells,play a critical role in maintaining immune tolerance and tissue homeostasis due to their potent immunosuppressive properties.Recent advances in research have highlighted the important therapeutic potential of Tregs in neurological diseases and tissue repair,emphasizing their multifaceted roles in immune regulation.This review aims to summarize and analyze the mechanisms of action and therapeutic potential of Tregs in relation to neurological diseases and neural regeneration.Beyond their classical immune-regulatory functions,emerging evidence points to non-immune mechanisms of regulatory T cells,particularly their interactions with stem cells and other non-immune cells.These interactions contribute to optimizing the repair microenvironment and promoting tissue repair and nerve regeneration,positioning non-immune pathways as a promising direction for future research.By modulating immune and non-immune cells,including neurons and glia within neural tissues,Tregs have demonstrated remarkable efficacy in enhancing regeneration in the central and peripheral nervous systems.Preclinical studies have revealed that Treg cells interact with neurons,glial cells,and other neural components to mitigate inflammatory damage and support functional recovery.Current mechanistic studies show that Tregs can significantly promote neural repair and functional recovery by regulating inflammatory responses and the local immune microenvironment.However,research on the mechanistic roles of regulatory T cells in other diseases remains limited,highlighting substantial gaps and opportunities for exploration in this field.Laboratory and clinical studies have further advanced the application of regulatory T cells.Technical advances have enabled efficient isolation,ex vivo expansion and functionalization,and adoptive transfer of regulatory T cells,with efficacy validated in animal models.Innovative strategies,including gene editing,cell-free technologies,biomaterial-based recruitment,and in situ delivery have expanded the therapeutic potential of regulatory T cells.Gene editing enables precise functional optimization,while biomaterial and in situ delivery technologies enhance their accumulation and efficacy at target sites.These advancements not only improve the immune-regulatory capacity of regulatory T cells but also significantly enhance their role in tissue repair.By leveraging the pivotal and diverse functions of Tregs in immune modulation and tissue repair,regulatory T cells–based therapies may lead to transformative breakthroughs in the treatment of neurological diseases.展开更多
Optimizing plant architecture for specific cultivation methods is essential for enhancing fruit productivity.Unlike indeterminate growth plants,the total productivity of determinate growth plants relies on cumulative ...Optimizing plant architecture for specific cultivation methods is essential for enhancing fruit productivity.Unlike indeterminate growth plants,the total productivity of determinate growth plants relies on cumulative fruit production and synchronized fruit ripening from both main and axillary shoots.Here,we focused on SlD14and SlMAX1,two key genes involved in the regulation of strigolactone(SL)signaling and biosynthesis,with the goal of maximizing yield and syn chronizing fruit ripening by fine-tuning axillary shoot growth.Using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)technology,we found that the sld14,slmax1,and sld14 slmax1mutant plants exhibited reduced plant height and increased axillary shoot proliferation compared to wild-type plants.However,these mutants showed reduced yield and delayed ripening,likely due to a source-sink imbalance caused by excessive axillary shoot development.A weak sld14 allele displayed a milder phenotype,maintaining total fruit yield and harvest index despite smaller individual fruit size.These findings indicate that allelic variation in SL-related genes can influence plant architecture and yield components.Our results suggest that weak or partial alleles may serve as promising targets for tailoring tomato architecture to space-limited cultivation systems.展开更多
High-quality silage is the cornerstone to sustainable livestock development and animal food production.As the core fermentation bacteria of silage,Lactobacillus directly regulates silage fermentation by producing lact...High-quality silage is the cornerstone to sustainable livestock development and animal food production.As the core fermentation bacteria of silage,Lactobacillus directly regulates silage fermentation by producing lactic acid,enzymes,and other bioactive molecules.However,traditional screening methods for functional strains are labor-intensive and time-consuming.Recent advances in synthetic biology,particularly the development of CRISPR-Cas genome editing technology,offer a revolutionary approach to designing Lactobacillus strains with customized traits.This review systematically reviewed the importance of silage in sustainable agricultural development and the limitations of current silage preparation and promotion.It also discussed the application of strain engineering approaches in optimizing the phenotypic performance of Lactobacillus for better silage.Building on this,we reviewed the research progress of CRISPR-Cas9 gene editing in Lactobacillus and discussed how to leverage its high efficiency and precision to optimize the strain's traits for improved silage quality and functionality.CRISPR-Cas9 toolkits are expected to achieve directed evolution of strain performance,ultimately yielding next-generation silage microbial inoculants with multiple functions,adaptability to multiple substrates,and eco-friendly characteristics.The use of such innovative biotechnologies would facilitate resource-efficient utilization,promote animal performance and health for sustainable development in livestock production.展开更多
This book is the proceedings of the NATO Advanced Science Institute“RemoteSensing of Processes Governing Energy and Water Cycles in the Climate System”,held inPln,Germany 1—12 May 1995.In this book,a total of 21 co...This book is the proceedings of the NATO Advanced Science Institute“RemoteSensing of Processes Governing Energy and Water Cycles in the Climate System”,held inPln,Germany 1—12 May 1995.In this book,a total of 21 conference papers are includedwith a special part for poster summaries.All the papers are modified or shorten versions展开更多
基金supported by Biological Breeding of Early Maturing and Disease Resistant Cotton Varieties (NO.2023ZD04041)the Project of China Agriculture Research System (Grant No. CARS-15-06)+2 种基金Natural Science Foundation of Henan Province (Grant No. 232300421041 and 222300420382)National Natural Science Foundation of China (Grant No. U21 A20213)the Central Public-interest Scientific Institution Basal Research Fund (Grant No. 1610162023017 and 1610162023028)。
文摘Background Cotton is an important crop providing the most natural fibers all over the world. The cotton genomics community has utilized whole genome sequencing data to construct an elite gene pool in which functional genes are related to agronomic traits. However, the functional validation of these genes is hindered by time-consuming and inefficient genetic transformation methods. Thus, establishing a transient transformation system of high efficiency is necessary for cotton genomics.Results To improve the efficiency of transient transformation, we used the protoplasts isolated from the etiolated cotyledon as recipient. The enzymatic digestion buffer comprised 1.5%(w/v) cellulase, 0.75%(w/v) macerozyme, and 1% hemicellulase, osmotically buffered with 0.4 mol·L^(-1) mannitol. After 5 h of dark incubation at 25℃, uniform cotton protoplasts were successfully isolated with a yield of 4.6 × 10^(6) protoplasts per gram(fresh weight) and 95% viability. We incubated 100 μL protoplasts(2.5 × 10^(5)·m L^(-1)) with 15 μg plasmid in the solution of 0.4 mol·L^(-1) mannitol and 40% PEG 4000 for 15 min, ultimately achieving an optimal transient transfection efficiency of 71.47%.Conclusions This transient system demonstrated effective utility in cellular biology research through successful applications in subcellular localization analyses, bimolecular fluorescence complementation(Bi FC) verification, and prime editing vector validation. Through systematic optimization, we established an efficient and expedited protoplast-based transient transformation system and successfully applied this platform to cotton functional genomics studies.
文摘Liver Transplantation (in Chinese), revised edition, edited by Professor Shu-Sen Zheng and prefaced by Professor Guo-Wei Sang, has just been published. The revised edition is mainly based on the clinical practice of the Liver Transplant Center, the First Affiliated Hospital, Zhejiang University and the experience
文摘This book was written by five authors. I had met two of them, the first and third author. The first author, Steven Wayne Lingafelter, is a research entomologist with the Systematic Entomology Laboratory, USDA, based at the Smithsonian Institution＇s National Museum of Natural History. He has specialized on longhomed woodboring beetles for almost three decades and currently specializes on the Neotropical fauna.
文摘Writing style is the essential issue even at the early stage the beginners who learnto read and write have to confront.From the part-Notes on reading and writing beforethe part of exercises of each lesson in English Book V-VⅢ we can come to see that the ed-itors attempt to mix the content(ideas)with the corresponding techniques.This is
基金financial support from the National Transgenic Key Project of the Chinese Natural Science Foundation (2016ZX08010-004,2016ZX08009001)the Beijing Natural Science Foundation (6162009)
文摘Genome editing is one of the most promising biotechnologies to improve crop performance.Common wheat is a staple food for mankind. In the past few decades both basic and applied research on common wheat has lagged behind other crop species due to its complex,polyploid genome and difficulties in genetic transformation. Recent breakthroughs in wheat transformation permit a revolution in wheat biotechnology. In this review, we summarize recent progress in wheat genetic transformation and its potential for wheat improvement. We then review recent progress in plant genome editing, which is now readily available in wheat. We also discuss measures to further increase transformation efficiency and potential applications of genome editing in wheat. We propose that, together with a high quality reference genome, the time for efficient genetic engineering and functionality studies in common wheat has arrived.
基金supported by Chinese Ministry of Agriculture and Rural Affairs (Grant No. 2018ZX0801003B)the National Transgenic Science and Technology Program (Grant No. 2016ZX08010002)
文摘Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR(TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35 S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.
基金Project supported by the National Natural Science Foundation of China(No.L1824000)
文摘The rapid developments of science and technology in China over recent decades, particularly in biomedical research, have brought forward serious challenges regarding ethical governance. Recently, Jian-kui HE, a Chinese scientist, claimed to have "created" the first gene-edited babies, designed to be naturally immune to the human immunodeficiency virus(HIV). The news immediately triggered widespread criticism, denouncement, and debate over the scientific and ethical legitimacy of HE’s genetic experiments. China’s guidelines and regulations have banned germline genome editing on human embryos for clinical use because of scientific and ethical concerns, in accordance with the international consensus. HE’s human experimentation has not only violated these Chinese regulations, but also breached other ethical and regulatory norms. These include questionable scientific value, unreasonable risk-benefit ratio, illegitimate ethics review, invalid informed consent, and regulatory misconduct. This series of ethical failings of HE and his team reveal the institutional failure of the current ethics governance system which largely depends on scientist’s self-regulation. The incident highlights the need for urgent improvement of ethics governance at all levels, the enforcement of technical and ethical guidelines, and the establishment of laws relating to such bioethical issues.
基金supported by the National Natural Science Foundation of China (31970574)。
文摘Animal models are extensively used in all aspects of biomedical research,with substantial contributions to our understanding of diseases,the development of pharmaceuticals,and the exploration of gene functions.The field of genome modification in rabbits has progressed slowly.However,recent advancements,particularly in CRISPR/Cas9-related technologies,have catalyzed the successful development of various genome-edited rabbit models to mimic diverse diseases,including cardiovascular disorders,immunodeficiencies,agingrelated ailments,neurological diseases,and ophthalmic pathologies.These models hold great promise in advancing biomedical research due to their closer physiological and biochemical resemblance to humans compared to mice.This review aims to summarize the novel gene-editing approaches currently available for rabbits and present the applications and prospects of such models in biomedicine,underscoring their impact and future potential in translational medicine.
基金supported by the fundings from the Region Pays de la Loire through Biogenouest,the IBiSA Program,Fondation Progreffethe French Government through the "Investissement d'avenir" program "TEFOR" project,managed by the National Research Agency(No.ANR-II-INSB-0014)the context of the "Investissement d'avenir" program LabEX IGO of the IHU-CESTI projects managed by the National Research Agency(Nos.ANR-11-LABX-001601 and ANR-10-IBHU-005,respectively)
文摘The recent emergence and application of engineered endonucleases have led to the development of genome editing tools capable of rapidly implementing various targeted genome editions in a wide range of species.Moreover,these novel tools have become easier to use and have resulted in a great increase of applications.Whilst gene knockout（KO） or knockin（KI） animal models are relatively easy to achieve,there is a bottleneck in the detection and analysis of these mutations.Although several methods exist to detect these targeted mutations,we developed a heteroduplex mobility assay on an automated microfluidic capillary electrophoresis system named HMA-CE in order to accelerate the genotyping process.The HMA-CE method uses a simple PCR amplification of genomic DNA（gDNA） followed by an automated capillary electrophoresis step which reveals a heteroduplexes（HD） signature for each mutation.This allows efficient discrimination of wild-type and genome-edited animals down to the single base pair level.
文摘Cotton,an important industrial crop cultivated in more than 70 countries,plays a major role in the livelihood of millions of farmers and industrialists.Cotton is mainly grown for its fiber,an economic component that can be differentiated from its epidermal cells in the outer integument of a developing seed.Fiber length,fiber strength,and fiber fineness are three main attributes that contribute to the quality of cotton fibers.Recent advancements in genomics have identified key genes,which are the most important factors that govern these three traits,can be introduced into cultivars of interest via gene editing,marker-assisted selection,and transgenics,thus the narrow genetic background of cotton can be addressed and its fiber quality traits can be enhanced.Over the past two decades,quantitative trait loci(QTLs)have been mapped for different fiber traits,approximately 1850 QTLs have been mapped for fiber length,fiber strength,and fineness among which a few genes have been edited for quality improvement in cotton.In this background,the current review covers the development and the factors that influence these traits,along with the reported genes,QTLs,and the edited genomes for trait improvement.
文摘The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;however,a few mismatches are tolerated by the Cas9 endonuclease activity.An accurate and careful in silico-based off-target prediction while target selection is preferred to address the issue.These predictions are based on a comprehensive set of selectable parameters.Therefore,we investigated the possible off-target prediction and their screening in StERF3 gene-edited potato plants while developing StERF3-loss-of-function mutants using CRISPR/Cas9 approach.The 201 off-targets for the selected targets of the StERF3 gene were predicted,and 79 werefiltered as potential off-targets.Of these 79,twenty-five off-targets showed scores with defined cut-off values<0.5 and were analyzed in Sterf3-edited potato plants compared to wild-type plants.No off-targeting was found to have occurred in edited plants.
基金supported by the National Natural Science Foundation of China(grant nos.32130031 and 31991201)and the China Postdoctoral Science Foundation(grant no.2021M702524).
文摘Dear Editor,Obtaining foreign DNA-free crops after genome editing is essential for agricultural applications,because of the current regulatory policies on gene-edited plants in various countries.However,it is usually a time-consuming and labor-consuming process to select and confirm foreign DNA-free offspring,especially in woody plants with long-term life cycles and crop species with complex genomes or T-DNA multisite insertions during genome editing.
基金Major Science and Technology Project of Yunnan Province(202102AA100054,202102AA310047)Basic Strengthening Program(2023-JCJQ-ZD-118-00/173)+2 种基金Xingdian Talent Support Program(XDYC-KJLJ-2022-0004)Major Science and Technology Project,Yunnan Province(202102AA310047,202102AA100054)National Key R&D Program of China(Grant No.2019YFA0110700).
文摘Xenotransplantation has entered the clinical phase in an effort to address the global organ shortage.However,recent clinical studies have revealed that current xenografts from gene-edited(GE)pigs still pose a risk of immune rejection and biosafety concerns.In this study,we successfully produced a large batch of 582 GE cloned(GEC)pigs with 10-(GTKO/CMAHKO/β4GalNT2KO/hCD46/hCD55/hCD59/hTBM/hCD39/hEPCR/hCD47)gene edits via gene editing and somatic cell cloning technologies,and successfully obtained the F1 generation.Phenotypic character ization of 10-GEC pigs revealed the deletion of three xenoantigens and the expression of seven human transgenes across various tissues.Digital droplet polymerase chain reaction and whole-genome sequencing revealed two cop ies of hCD46/hCD55/hCD59/hTBM/hCD39 and one copy of hEPCR/hCD47 in the pig genome with minimal off-target effects or damage to the porcine functional genes.The validation results showed that 10-GEC pigs could effectively inhibit attacks from human antibodies,complement and macrophages on porcine endothelial cells,and alleviated coagulation abnormalities between pigs and humans.Large-scale screening of pathogens revealed no evidence of 47 pathogens,including cytomegalovirus,in our 10-GEC pigs.Kidney,heart and liver xenografts from these 10-GEC pigs were transplanted into nonhuman primates(NHPs),which worked normally without hyperacute rejection(HAR).Among NHPs,the heart and liver orthotopic transplant recipients survived for 3 and 4 days,respectively,while the two kidney transplant recipients survived for 23 and 16 days,respectively.Pathological analysis showed interstitial hemorrhage and fibrosis,cellular hyperplasia with minor antibodies and complement deposition,but significantly reduced infiltration of CD68^(+)macrophages in 10-GEC pig kidney xenografts.In summary,we success fully produced specific pathogen-free 10-GEC donor pigs that resulted in effective mitigation of immune rejection upon multiorgan transplantation to NHPs.
基金supported by grants from the National Key Research and Development Program of China(2023YFD1202905)the National Natural Science Foundation of China(32272629).
文摘Dear Editor,Genome editing tools are leading a revolution in plant breeding.In particular,prime editors(PEs)can install all types of base changes and small insertions/deletions at precise positions in plant genomes(Anzalone et al.,2019).PEs are by far the most powerful approach for improving traits conferred by gain-of-function point mutations.Early versions of PEs suffered from low editing efficiency,but the latest PEs can perform edits at a much higher efficiency thanks to the extensive efforts of re-searchers from around the world.Most modifications to improve PE efficiency have focused on the optimization of PE protein components and structure.
基金the National Major Transgenic Breeding Project(2008ZX08006-003,2011ZX08006-003,2013ZX08006-003,2014ZX08006-003,and 2016ZX08006-001)the Na-tional Key Basic Research Development Plan(2015CB943100)+1 种基金the Key Projects of the National Natural Science Foundation of China(30830080 and 31330074)China Postdoctoral Foundation Project(2018M631648)。
文摘Beef and mutton production has been aided by breeding to integrate allelic diversity for myostatin(MSTN),but a lack of diversity in the MSTN germplasm has limited similar advances in pig farming.Moreover,insurmountable challenges with congenital lameness and a dearth of data about the impacts of feed conversion,reproduction,and meat quality in MSTN-edited pigs have also currently blocked progress.Here,in a largest-to-date evaluation of multiple MSTN-edited pig populations,we demonstrated a practical alternative edit-site-based solution that overcomes the major production obstacle of hindlimb weakness.We also provide long-term and multidomain datasets for multiple breeds that illustrate how MSTN-editing can sustainably increase the yields of breed-specific lean meat and the levels of desirable lipids without deleteriously affecting feed-conversion rates or litter size.Apart from establishing a new benchmark for the data scale and quality of genome-edited animal production,our study specifically illustrates how gene-editing site selection profoundly impacts the phenotypic outcomes in diverse genetic back-grounds.
基金National Science Foundation grant no. IOS-1546625 (GBM and ZF).
文摘Recent advances in genome editing technologies, particularly CRISPR/Cas, enable the alteration of DNA sequences to produce deletions, insertions, and substitutions in genes (Jaganathan et al., 2018), as well as large or entire chromosome deletions in the genomes of plants and animals (Zhou et al., 2014;Adikusuma et al., 2017).
基金supported by the National Natural Science Foundation of China,Nos.32271389,31900987(both to PY)the Natural Science Foundation of Jiangsu Province,No.BK20230608(to JJ)。
文摘Regulatory T cells,a subset of CD4^(+)T cells,play a critical role in maintaining immune tolerance and tissue homeostasis due to their potent immunosuppressive properties.Recent advances in research have highlighted the important therapeutic potential of Tregs in neurological diseases and tissue repair,emphasizing their multifaceted roles in immune regulation.This review aims to summarize and analyze the mechanisms of action and therapeutic potential of Tregs in relation to neurological diseases and neural regeneration.Beyond their classical immune-regulatory functions,emerging evidence points to non-immune mechanisms of regulatory T cells,particularly their interactions with stem cells and other non-immune cells.These interactions contribute to optimizing the repair microenvironment and promoting tissue repair and nerve regeneration,positioning non-immune pathways as a promising direction for future research.By modulating immune and non-immune cells,including neurons and glia within neural tissues,Tregs have demonstrated remarkable efficacy in enhancing regeneration in the central and peripheral nervous systems.Preclinical studies have revealed that Treg cells interact with neurons,glial cells,and other neural components to mitigate inflammatory damage and support functional recovery.Current mechanistic studies show that Tregs can significantly promote neural repair and functional recovery by regulating inflammatory responses and the local immune microenvironment.However,research on the mechanistic roles of regulatory T cells in other diseases remains limited,highlighting substantial gaps and opportunities for exploration in this field.Laboratory and clinical studies have further advanced the application of regulatory T cells.Technical advances have enabled efficient isolation,ex vivo expansion and functionalization,and adoptive transfer of regulatory T cells,with efficacy validated in animal models.Innovative strategies,including gene editing,cell-free technologies,biomaterial-based recruitment,and in situ delivery have expanded the therapeutic potential of regulatory T cells.Gene editing enables precise functional optimization,while biomaterial and in situ delivery technologies enhance their accumulation and efficacy at target sites.These advancements not only improve the immune-regulatory capacity of regulatory T cells but also significantly enhance their role in tissue repair.By leveraging the pivotal and diverse functions of Tregs in immune modulation and tissue repair,regulatory T cells–based therapies may lead to transformative breakthroughs in the treatment of neurological diseases.
基金funded by the National Research Foundation of Korea(NRF)grant from the Ministry of Science and ICT(MSIT),Republic of Korea(Nos.RS-2024-00407469 and RS-2025-00517964)the BK21 FOUR program of Graduate School,Kyung Hee University(GS-1-JO-NON-20240417)。
文摘Optimizing plant architecture for specific cultivation methods is essential for enhancing fruit productivity.Unlike indeterminate growth plants,the total productivity of determinate growth plants relies on cumulative fruit production and synchronized fruit ripening from both main and axillary shoots.Here,we focused on SlD14and SlMAX1,two key genes involved in the regulation of strigolactone(SL)signaling and biosynthesis,with the goal of maximizing yield and syn chronizing fruit ripening by fine-tuning axillary shoot growth.Using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)technology,we found that the sld14,slmax1,and sld14 slmax1mutant plants exhibited reduced plant height and increased axillary shoot proliferation compared to wild-type plants.However,these mutants showed reduced yield and delayed ripening,likely due to a source-sink imbalance caused by excessive axillary shoot development.A weak sld14 allele displayed a milder phenotype,maintaining total fruit yield and harvest index despite smaller individual fruit size.These findings indicate that allelic variation in SL-related genes can influence plant architecture and yield components.Our results suggest that weak or partial alleles may serve as promising targets for tailoring tomato architecture to space-limited cultivation systems.
基金supported by the National Nature Science Foundation of China(No.U20A2002)。
文摘High-quality silage is the cornerstone to sustainable livestock development and animal food production.As the core fermentation bacteria of silage,Lactobacillus directly regulates silage fermentation by producing lactic acid,enzymes,and other bioactive molecules.However,traditional screening methods for functional strains are labor-intensive and time-consuming.Recent advances in synthetic biology,particularly the development of CRISPR-Cas genome editing technology,offer a revolutionary approach to designing Lactobacillus strains with customized traits.This review systematically reviewed the importance of silage in sustainable agricultural development and the limitations of current silage preparation and promotion.It also discussed the application of strain engineering approaches in optimizing the phenotypic performance of Lactobacillus for better silage.Building on this,we reviewed the research progress of CRISPR-Cas9 gene editing in Lactobacillus and discussed how to leverage its high efficiency and precision to optimize the strain's traits for improved silage quality and functionality.CRISPR-Cas9 toolkits are expected to achieve directed evolution of strain performance,ultimately yielding next-generation silage microbial inoculants with multiple functions,adaptability to multiple substrates,and eco-friendly characteristics.The use of such innovative biotechnologies would facilitate resource-efficient utilization,promote animal performance and health for sustainable development in livestock production.
文摘This book is the proceedings of the NATO Advanced Science Institute“RemoteSensing of Processes Governing Energy and Water Cycles in the Climate System”,held inPln,Germany 1—12 May 1995.In this book,a total of 21 conference papers are includedwith a special part for poster summaries.All the papers are modified or shorten versions
