Since the discovery of the double helix structure of deoxyribonucleic acid(DNA),significant progress has been made in engineering technologies such as gene analysis and editing,greatly promoting the development of bio...Since the discovery of the double helix structure of deoxyribonucleic acid(DNA),significant progress has been made in engineering technologies such as gene analysis and editing,greatly promoting the development of biomedical research and clinical applications.In recent years,the rapid development of genome-editing technologies,especially the clustered regularly interspaced short palindromic repeats-associated protein 9(CRISPR-Cas9)system,have brought unprecedented opportunities for biomedical research and clinical applications.However,with the widespread application of genome-editing technologies,biosafety issues have gradually attracted the attention of the scientific community and the public.Potential risks such as off-target effects,genomic instability,and ethical and legal issues need to be taken seriously,and their safety and effectiveness in clinical applications still require further verification.This review summarizes the current status and potential risks of gene editing technologies in the medical field and discusses how to ensure its safe application through policies,regulations,and technical means.Through in-depth exploration of these issues,we hope to provide a comprehensive perspective for the scientific community,policy makers,and the public to promote the safe and responsible application of genome-editing technologies.展开更多
To explore how rice(Oryza sativa L.) can be safely produced in Cd-polluted soil, OsLCT1 and OsNramp5 mutant lines were generated by CRISPR/Cas9-mediated mutagenesis. One of OsLCT1 mutant(lct1×1) and two of OsNram...To explore how rice(Oryza sativa L.) can be safely produced in Cd-polluted soil, OsLCT1 and OsNramp5 mutant lines were generated by CRISPR/Cas9-mediated mutagenesis. One of OsLCT1 mutant(lct1×1) and two of OsNramp5 mutants(nramp5×7 and nramp5×9) were evaluated for grain Cd accumulation and agronomic performances. In paddy field soil containing approximately 0.9 mg/kg Cd, lct1×1 grains contained approximately 40%(0.17 mg/kg) of the Cd concentration of the wild type parental line, less than the China National Food Safety Standard(0.20 mg/kg). Both OsNramp5 mutants showed low grain Cd accumulation(< 0.06 mg/kg) in the paddy(approximately 0.9 mg/kg Cd) or in pots in soil spiked with 2 mg/kg Cd. However, only nramp5×7 showed normal growth and yield, whereas the growth of nramp5×9 was severely impaired. The study showed that lct1×1 could be used to produce rice grains safe for human consumption in lightly contaminated paddy soils and nramp5×7 used in soils contaminated by much higher levels of Cd.展开更多
Chronic hepatitis B infection is caused by hepatitis B virus(HBV) and a total cure is yet to be achieved. The viral covalently closed circular DNA(ccc DNA) is the key to establish a persistent infection within hepatoc...Chronic hepatitis B infection is caused by hepatitis B virus(HBV) and a total cure is yet to be achieved. The viral covalently closed circular DNA(ccc DNA) is the key to establish a persistent infection within hepatocytes. Current antiviral strategies have no effect on the pre-existing ccc DNA reservoir. Therefore, the study of the molecular mechanism of ccc DNA formation is becoming a major focus of HBV research. This review summarizes the current advances in ccc DNA molecular biology and the latest studies on the elimination or inactivation of ccc DNA, including three major areas:(1) epigenetic regulation of ccc DNA by HBV X protein,(2) immune-mediated degradation,and(3) genome-editing nucleases. All these aspects provide clues on how to finally attain a cure for chronic hepatitis B infection.展开更多
Crop breeding requires a balance of tradeoffs among key agronomic traits caused by gene pleiotropy.The molecular manipulation of genes can effectively improve target traits,but this may not reduce gene pleiotropy,pote...Crop breeding requires a balance of tradeoffs among key agronomic traits caused by gene pleiotropy.The molecular manipulation of genes can effectively improve target traits,but this may not reduce gene pleiotropy,potentially leading to undesirable traits or even lethal conditions.However,molecular editing of cis-regulatory elements(CREs)of target genes may facilitate the dissection of gene pleiotropy to fine-tune gene expression.In this study,we developed a pipeline,in potato,which employs open chromatin to predict candidate CREs,along with both transient and genetic assays to validate the function of CREs and CRISPR/Cas9 to edit candidate CREs.We used StCDF1 as an example,a key gene for potato tuberization and identified a 288 bp-core promoter region,which showed photoperiodic inducibility.A homozygous CRISPR/Cas9-editing line was established,with two deletions in the core promoter,which displayed a reduced expression level,resulting in late tuberization under both longday and short-day conditions.This pipeline provides an alternative pathway to improve a specific trait with limited downside on other phenotypes.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
基金supported by The National Key Research and Development Program(2022YFC2603500,2022YFC2603505)Beijing Hospitals Authority Clinical medicine Development of Special Funding(XMLX 202127)+5 种基金The Digestive Medical Coordinated Development Center of Beijing Hospitals Authority(XXZ0302)The Capital Health Research and Development of Special Public Health Project(2022-1-2172)Beijing Municipal Health Commission High-level Public Health Technical Personnel Construction Project(discipline leader-03-26)Beijing Hospitals Authority Clinical Medicine Development of Special FundingBeijing Hospitals Authority“Peak”Talent Training Program(DFL20241803)The National Key Research and Development Program(2023YFC2306901,2023YFC2308105).
文摘Since the discovery of the double helix structure of deoxyribonucleic acid(DNA),significant progress has been made in engineering technologies such as gene analysis and editing,greatly promoting the development of biomedical research and clinical applications.In recent years,the rapid development of genome-editing technologies,especially the clustered regularly interspaced short palindromic repeats-associated protein 9(CRISPR-Cas9)system,have brought unprecedented opportunities for biomedical research and clinical applications.However,with the widespread application of genome-editing technologies,biosafety issues have gradually attracted the attention of the scientific community and the public.Potential risks such as off-target effects,genomic instability,and ethical and legal issues need to be taken seriously,and their safety and effectiveness in clinical applications still require further verification.This review summarizes the current status and potential risks of gene editing technologies in the medical field and discusses how to ensure its safe application through policies,regulations,and technical means.Through in-depth exploration of these issues,we hope to provide a comprehensive perspective for the scientific community,policy makers,and the public to promote the safe and responsible application of genome-editing technologies.
基金supported by the Zhejiang Provincial S & T Project on Breeding of Agricultural (Food) Crops (Grant No. 2016C02050-2)
文摘To explore how rice(Oryza sativa L.) can be safely produced in Cd-polluted soil, OsLCT1 and OsNramp5 mutant lines were generated by CRISPR/Cas9-mediated mutagenesis. One of OsLCT1 mutant(lct1×1) and two of OsNramp5 mutants(nramp5×7 and nramp5×9) were evaluated for grain Cd accumulation and agronomic performances. In paddy field soil containing approximately 0.9 mg/kg Cd, lct1×1 grains contained approximately 40%(0.17 mg/kg) of the Cd concentration of the wild type parental line, less than the China National Food Safety Standard(0.20 mg/kg). Both OsNramp5 mutants showed low grain Cd accumulation(< 0.06 mg/kg) in the paddy(approximately 0.9 mg/kg Cd) or in pots in soil spiked with 2 mg/kg Cd. However, only nramp5×7 showed normal growth and yield, whereas the growth of nramp5×9 was severely impaired. The study showed that lct1×1 could be used to produce rice grains safe for human consumption in lightly contaminated paddy soils and nramp5×7 used in soils contaminated by much higher levels of Cd.
基金supported by the Key Project of Hubei Province Natural Science Foundation(2014CFA075)the National Natural Science Foundation of China(31400153)the Applied Basic Research Program(2015060101010033),Wuhan,China
文摘Chronic hepatitis B infection is caused by hepatitis B virus(HBV) and a total cure is yet to be achieved. The viral covalently closed circular DNA(ccc DNA) is the key to establish a persistent infection within hepatocytes. Current antiviral strategies have no effect on the pre-existing ccc DNA reservoir. Therefore, the study of the molecular mechanism of ccc DNA formation is becoming a major focus of HBV research. This review summarizes the current advances in ccc DNA molecular biology and the latest studies on the elimination or inactivation of ccc DNA, including three major areas:(1) epigenetic regulation of ccc DNA by HBV X protein,(2) immune-mediated degradation,and(3) genome-editing nucleases. All these aspects provide clues on how to finally attain a cure for chronic hepatitis B infection.
基金supported by grants from the National Natural Science Foundation of China(32170573)Sichuan Science and Technology Program(2021YFYZ0019,2023ZYD0056).
文摘Crop breeding requires a balance of tradeoffs among key agronomic traits caused by gene pleiotropy.The molecular manipulation of genes can effectively improve target traits,but this may not reduce gene pleiotropy,potentially leading to undesirable traits or even lethal conditions.However,molecular editing of cis-regulatory elements(CREs)of target genes may facilitate the dissection of gene pleiotropy to fine-tune gene expression.In this study,we developed a pipeline,in potato,which employs open chromatin to predict candidate CREs,along with both transient and genetic assays to validate the function of CREs and CRISPR/Cas9 to edit candidate CREs.We used StCDF1 as an example,a key gene for potato tuberization and identified a 288 bp-core promoter region,which showed photoperiodic inducibility.A homozygous CRISPR/Cas9-editing line was established,with two deletions in the core promoter,which displayed a reduced expression level,resulting in late tuberization under both longday and short-day conditions.This pipeline provides an alternative pathway to improve a specific trait with limited downside on other phenotypes.
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).
