Objective:This randomized controlled trial aimed to assess the impact of a targeted educational intervention on the awareness and practice of genetic screening and counseling among young adults in Calabar Municipality...Objective:This randomized controlled trial aimed to assess the impact of a targeted educational intervention on the awareness and practice of genetic screening and counseling among young adults in Calabar Municipality,Cross River State.Materials and Methods:Participants(aged 18-45)were randomly assigned to either an intervention group receiving structured health education or a control group receiving general health information.Stratified randomization was used between the groups.A sample size of 340 participants was recruited to detect a 20%difference in outcomes with 80%power.Data were collected using prevalidated questionnaires at baseline,immediately after the intervention,and at a 3-month follow-up.The intervention consisted of three weekly 90-min educational sessions covering genetics,the benefits of screening,and practical guidance on accessing genetic services.The primary outcomes were changes in awareness and practices related to genetic screening,with secondary outcomes focusing on attitudes and intentions toward genetic counseling.Results:Findings revealed that awareness of genetic screening was higher in the intervention group,with 65.9%of participants aware of early detection,compared to 59.4%in the control group,although the difference was not statistically significant(P=0.23).In terms of practice,42.9%of the intervention group and 40.0%of the control group engaged in genetic screening programs,with no significant difference(P=0.57).Logistic regression analysis highlighted that age,educational level,and knowledge of teratogens were significant predictors of genetic screening awareness.Participants aged 36 years and above were 1.52 times more likely to be aware(odds ratio[OR]=1.52,P=0.003),and those with tertiary education had nearly double the likelihood of awareness(OR=1.96,P<0.001).Conclusion:The study underscores the importance of targeted education in improving genetic screening awareness.展开更多
Liver cancer,primarily hepatocellular carcinoma,remains a global health challenge with rising incidence and limited therapeutic options.Genetic factors play a pivotal role in the development and progression of liver c...Liver cancer,primarily hepatocellular carcinoma,remains a global health challenge with rising incidence and limited therapeutic options.Genetic factors play a pivotal role in the development and progression of liver cancer.This state-of-the-art paper provides a comprehensive review of the current landscape of genetic screening strategies for liver cancer.We discuss the genetic underpinnings of liver cancer,emphasizing the critical role of risk-associated genetic variants,somatic mutations,and epigenetic alterations.We also explore the intricate interplay between environmental factors and genetics,highlighting how genetic screening can aid in risk stratification and early detection via using liquid biopsy,and advancements in high-throughput sequencing technologies.By synthesizing the latest research findings,we aim to provide a comprehensive overview of the state-of-the-art genetic screening methods for liver cancer,shedding light on their potential to revolutionize early detection,risk assessment,and targeted therapies in the fight against this devastating disease.展开更多
Objective:To detect common chromosomal aneuploidy variations in embryos from couples undergoing assisted reproductive technology and preimplantation genetic screening and their possible associations with embryo qualit...Objective:To detect common chromosomal aneuploidy variations in embryos from couples undergoing assisted reproductive technology and preimplantation genetic screening and their possible associations with embryo quality.Methods:In this study,359 embryos from 62 couples were screened for chromosomes 13,21,18,X,and Y by fluorescence insitu hybridization.For biopsy of blastomere,a laser was used to remove a significantly smaller portion of the zona pellucida.One blastomere was gently biopsied by an aspiration pipette through the hole.After biopsy,the embryo was immediately returned to the embryo scope until transfer.Embryo integrity and blastocyst formation were assessed on day 5.Results:Totally,282 embryos from 62 couples were evaluated.The chromosomes were normal in 199(70.57%)embryos and abnormal in 83(29.43%)embryos.There was no significant association between the quality of embryos and numerical chromosomal abnormality(P=0.67).Conclusions:Embryo quality is not significantly correlated with its genetic status.Hence,the quality of embryos determined by morphological parameters is not an appropriate method for choosing embryos without these abnormalities.展开更多
BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromi...BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens.AIM To derive human haploid neural stem cells(haNSCs)to carry out lineage-specific screens.METHODS Human haNSCs were differentiated from human extended haESCs with the help of Y27632(ROCK signaling pathway inhibitor)and a series of cytokines to reduce diploidization.Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency.Global gene expression analysis was conducted to compare haNSCs with diploid NSCs and haESCs.Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs.Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools.RESULTS Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies,a higher efficiency in neural differentiation,a higher cell survival ratio and higher stability in haploidy maintenance.These characteristics effectively facilitated the derivation of human haNSCs.These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro.After PiggyBac transfection,there were multiple insertion sites in the human haNSCs’genome,and the insertion sites were evenly spread across all chromosomes.In addition,after the cells were treated with manganese,we were able to generate a list of manganese-induced toxicity genes,demonstrating their utility as genetic screening tools.CONCLUSION This is the first report of a generated human haploid somatic cell line with a complete genome,proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.展开更多
Leukemia, like many other cancers, is thought to arise from a small population of stem cells that have the capacity to self-renewal extensively and to initiate, sustain or regenerate the disease. Elimination of the le...Leukemia, like many other cancers, is thought to arise from a small population of stem cells that have the capacity to self-renewal extensively and to initiate, sustain or regenerate the disease. Elimination of the leukemia stem cells （LSCs） will likely be essential, and probably sufficient, for curing this disease.Recent studies have shown that LSCs can be derived from early hematopoietic progenitors as well as more differentiated derivatives; the key feature being these cells have acquired an increased proliferative capacity and the ability to self-renew extensively. Genes that make this possible are attractive drug targets for treating leukemia. In our laboratory we have developed a novel in vitro genetic screen that uses retroviral insertional mutagenesis as a tool for identifying genes that are able to convert both normal hematoooietic progenitors and committed mveloid progenitor cells into cells that resemble LSCs.展开更多
Objectives The objective of this study was to evaluate the role of genetic screening in a Chinese woman of childbearing age with hypertrophic cardiomyopathy.Methods and Results One 39 year-old woman with presyncope fo...Objectives The objective of this study was to evaluate the role of genetic screening in a Chinese woman of childbearing age with hypertrophic cardiomyopathy.Methods and Results One 39 year-old woman with presyncope for 10 years was admitted.Both of her father and her son died of sudden death and she strongly desire to get another baby. A series of clinical and laboratory studies were performed. Hypertrophic cardiomyopathy was diagnosed finally and implantable Cardioverter Defibrillator was implanted to prevent sudden cardiac death for her.Genomic DNA was isolated and the most common causal genes for hypertrophic cardiomyopathy were screened.A known pathogenetic heterozygous mutation c.700 g】a(p.Argl86Gln) in TNNI3 was found,which was not found in 100 normal control individuals matched for age,sex and geographical region.Because 50%probability of the pathogenetic mutation is inherited to the offsprings,she will get a healthy baby in vitro fertilization which the egg comes from a healthy female donor to prevent from the inherited HCM.Conclusions We found a pathogenetic TNNI3 mutation in a Chinese woman of childbearing age with hypertrophic cardiomyopathy.The genetic screening definite DNA-based diagnosis and help to design a healthy fertilization in vitro for female with hypertrophic cardiomyopathy.展开更多
CRISPR technologies have revolutionized research areas ranging from fundamental science to translational medicine.CRISPR-based genetic screens offer a powerful platform for unbiased screening in various fields,such as...CRISPR technologies have revolutionized research areas ranging from fundamental science to translational medicine.CRISPR-based genetic screens offer a powerful platform for unbiased screening in various fields,such as cancer immunology.Immune checkpoint blockade(ICB)therapy has been shown to strongly affect cancer treatment.However,the currently available ICBs are limited and do not work in all cancer patients.Pooled CRISPR screens enable the identification of previously unknown immune regulators that can regulate T-cell activation,cytotoxicity,persistence,infiltration into tumors,cytokine secretion,memory formation,T-cell metabolism,and CD4^(+)T-cell differentiation.These novel targets can be developed as new immunotherapies or used with the current ICBs as new combination therapies that may yield synergistic efficacy.Here,we review the progress made in the development of CRISPR technologies,particularly technological advances in CRISPR screens and their application in novel target identification for immunotherapy.展开更多
Objective To screen and identify genetic loci affecting the active zone formation in C. elegans. Methods A SYD-2：：GFP reporter was constructed and used as an active zone marker for forward genetic screen to identify...Objective To screen and identify genetic loci affecting the active zone formation in C. elegans. Methods A SYD-2：：GFP reporter was constructed and used as an active zone marker for forward genetic screen to identify genetic loci affecting the active zone formation. Results Eight isolated mutant alleles were characterized from 15,000 haploid genomes. The SYD-2：：GFP phenotypes of these mutants are mainly reflected as the changes of number, morphology, distribution of puncta and the gaps appearance. Some mutants also exhibit visible behavioral or physical phenotypes, and aldicarb resistant or sensitive phenotypes. Conclusion These mutants provide the opportunity for further systematic research on the active zone formation and the neurotransmission.展开更多
The Saudi genome project started in 2013 with a great hope to improve medical care and disease prevention.Among the genes are those related to nutrition and fitness that can optimize an individual’s lifestyle.Our aim...The Saudi genome project started in 2013 with a great hope to improve medical care and disease prevention.Among the genes are those related to nutrition and fitness that can optimize an individual’s lifestyle.Our aim was to review the knowledge and acceptance of nutrition and fitness genetic testing to enhance the quality of life among the population of Saudi Arabia.For the study an electronic questionnaire consisting of 27 questions was prepared,and it was answered by 302 respondents.The respondents’demographics showed about 50% of respondents were aged 18-25 years and about 50% of respondents were aged 26-60 years.More than 50% of respondents were interested in having a genetic test to enhance their health,while 40% were interested in having a genetic test to enhance their fitness.Less than 50% of respondents had an understanding of the effects of coffee,macronutrition and micronutrition,elements,and enzyme activity.These results represented a contribution to the discussion on the relevance of genetic testing validity and acceptance among the population of Saudi Arabia.The results might help in producing specific guidelines on genetic testing and genomic analysis and help in the implementation of fitness and future health plans in cooperation with Saudi genome projects.Future study will focus on population structure and genetic frequency related to specific diets or fitness.展开更多
The utilisation of polygenic scoring models may enhance the clinician’s ability to risk stratify an inflammatory bowel disease patient’s individual risk for venous thromboembolism(VTE)and guide the appropriate usage...The utilisation of polygenic scoring models may enhance the clinician’s ability to risk stratify an inflammatory bowel disease patient’s individual risk for venous thromboembolism(VTE)and guide the appropriate usage of VTE thromboprophylaxis,yet there is a need to validate such models in ethnically diverse populations.展开更多
The advancement of Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)gene editing technology has revolutionized the comprehension of human genome,propelling molecular and cellular biology research into ...The advancement of Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)gene editing technology has revolutionized the comprehension of human genome,propelling molecular and cellular biology research into unexplored realms and accelerating progress in life sciences and medicine.CRISPR-based gene screening,recognized for its efficiency and practicality,is widely utilized across diverse biological fields.Aging is a multifaceted process governed by a myriad of genetic and epigenetic factors.Unraveling the genes regulating aging holds promise for understanding this intricate phenomenon and devising strategies for its assessment and intervention.This review provides a comprehensive overview of the progress in CRISPR screening and its applications in aging research,while also offering insights into future directions.CRISPR-based genetic-manipulation tools are positioned as indispensable instruments for mitigating aging and managing age-related diseases.展开更多
Charcot-Marie-Tooth （CMT） disease is the most common hereditary neuropathy, with a population prevalence of 1 in 2500. CMT disease type 1A （CMT1A）, accounting for ~70% of CMT1 cases and ~ 50% of all CMT cases, is ...Charcot-Marie-Tooth （CMT） disease is the most common hereditary neuropathy, with a population prevalence of 1 in 2500. CMT disease type 1A （CMT1A）, accounting for ~70% of CMT1 cases and ~ 50% of all CMT cases, is transmitted in an autosomal dominant manner. CMT1A maps to chromo- some 17pl 1.2 and is caused, in the majority of cases, by a 1.4- Mb tandem duplication that includes the peripheral myelin protein22 （PMP22） gene （Li et al., 2013）. The disease usually presents in the first 20 years of age, causing difficulty in walking or running, distal symmetrical muscle weakness and wasting, and sensory loss （van Paassen et al., 2014）.展开更多
The rapidly evolving environment of assisted reproductive technology(ART)requires consideration of how new innovations are reshaping clinical practice as much as research.In particular,there are three key areas that,w...The rapidly evolving environment of assisted reproductive technology(ART)requires consideration of how new innovations are reshaping clinical practice as much as research.In particular,there are three key areas that,while full of promise,also present significant challenges:the use of artificial intelligence(AI)in embryo selection,the impact of personalized medicine on ART success rates,and the ethical considerations of genetic screening of embryos[1].This letter is meant to provoke further discussion and highlight the need for harmonized global guidelines as these advances continue to reshape the reproductive medicine environment.展开更多
Objective:Lynch syndrome(LS)increases the risk of various cancers,including colorectal cancer,endometrial cancer and gastric cancer(GC).The incidence of LS among microsatellite instability-high(MSI-H)GC and their asso...Objective:Lynch syndrome(LS)increases the risk of various cancers,including colorectal cancer,endometrial cancer and gastric cancer(GC).The incidence of LS among microsatellite instability-high(MSI-H)GC and their association in South Korea remains underexplored.This study investigates LS-associated pathogenic germline variants in MSI-H GC patients using whole-exome sequencing(WES)on normal tissues.Methods:This retrospective study included patients who underwent gastrectomy for GC at Soonchunhyang University Bucheon and Cheonan Hospitals from January 2011 to October 2023.Among 1,537 patients screened for MSI status,127(8.3%)were identified as MSI-H.WES was performed on normal tissues from 123 patients.Pathogenic/likely pathogenic(P/LP)variants in mismatch repair(MMR)genes were identified using in silico models and protein loss assessments in corresponding tumor tissues.Results:Of the 127 MSI-H GC cases,characteristics aligned with typical MSI-H GC.The average age was70.02 years,with 98(77.2%)located in the lower body and 81(63.8%)of the intestinal type.All five MSI markers were positive in 46.5%of cases,whereas four markers were positive in 27.6%.Of the MSI-H GCs,10 LS candidates were identified.Three patients had known P/LP variants[MLH1(c.1758dup),MSH6(c.3261dup),MSH2(c.1241T>C)].Seven patients had variants of unknown significance(VUS)in MMR genes.Six(4.9%)patients were identified as having LS or possible LS,including one patient with the MLH1(c.1153C>T)variant previously classified as VUS but now considered LS-associated.Conclusions:This large-scale screening for LS in MSI-H GC patients using retrospective samples confirmed the lower incidence of LS than those of colorectal or endometrial cancer and GC patients in Western countries,emphasizing the need for clinical consideration in managing MSI-H GC patients.展开更多
Trophoblast stem cells(TSCs)are placental progenitors derived from the trophectoderm or the extraembryonic ectoderm.Recently,human TSCs were established via a specific culture system[1].However,the process by which hu...Trophoblast stem cells(TSCs)are placental progenitors derived from the trophectoderm or the extraembryonic ectoderm.Recently,human TSCs were established via a specific culture system[1].However,the process by which human TSCs further differentiate into terminal trophoblasts that mimic placental development remains obscure and interesting.Hence,the development of a suitable celltool to robustly study the mechanism underlying the self-renewal or differentiation of human TSCs is urgently needed.Mammalian haploid cells resemble their diploid counterparts but have a single genome,which is widely used in the genetic screening of many biological processes.However,no haploid cells for an extraembryonic lineage in humans have been derived.展开更多
The breeding of herbicide-resistant wheat varieties has helped control weeds in wheat fields economically and effectively.Imidazolinone (IMI) herbicides are popular as they have low toxicity in mammals,are effective a...The breeding of herbicide-resistant wheat varieties has helped control weeds in wheat fields economically and effectively.Imidazolinone (IMI) herbicides are popular as they have low toxicity in mammals,are effective at small doses,and exhibit broad-spectrum herbicidal action in the field.Therefore,the isolation and genetic and molecular characterization of IMI-resistant wheat mutants will enhance weed management in wheat fields.In the present study,352 IMI-resistant plants were isolated by genetic screening from a mutant pool prepared by EMS-based random mutagenesis.Cloning of the mutated genes from the IMI-resistant plants indicated that ten taals alleles had been isolated,and mutation in one of three Ta ALS homolog genes conferred IMI resistance,and such a mutation is a dominant trait.Further analysis showed that taals-d exhibited the greatest IMI resistance,whereas taals-b exhibited the weakest resistance to IMI among three homologous taals mutants.In terms of IMI resistance,the taals triple mutant was stronger than the taals double mutants,and the taals double mutants were stronger than the single mutants,indicating a dose-dependent effect of the Ta ALS mutation on IMI resistance in wheat.Biochemical analysis indicated that the mutation in Ta ALS increased the tolerance of Ta ALS to inhibition by IMI.Our work details the genetic and molecular characterization of als wheat mutants,provides a foundation for understanding IMI resistance and breeding wheat varieties with herbicide resistance,and indicates that genetic screening using a mutagenized pool is an effective and important means of breeding crops with additional desired agricultural traits.展开更多
This paper offers a general review and comparative analysis of various types of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technologies. It evaluates the strengths and weaknesses of these techn...This paper offers a general review and comparative analysis of various types of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technologies. It evaluates the strengths and weaknesses of these technologies to identify the optimal approach for conducting genetic screens. Through an extensive literature review, this paper examines CRISPR nuclease, CRISPR activation (CRISPRa), and CRISPR interference (CRISPRi) screens. This study concludes that CRISPRa and CRISPRi are more advantageous due to their use of deactivated Cas9 proteins that only over-express or deactivate genes rather than irreversibly breaking genes like CRISPRn. Notably, CRISPRa is unique in its ability to over-express genes, while the other two technologies deactivate genes. Future studies may focus on inducing multiple mutations simultaneously—both gain-of-function and gene knockout—to carry out a more complete screen that can test the combinatorial effect of genes. Likewise, targeting both exons and introns can offer a more thorough understanding of a specific phenotype.展开更多
A wing specific F 1 genetic screen was carried out using the powerful Drosophila genetic system, combined with yeast FRT/FLP and GAL4/UAS system. Form the wing phenotypes and germline clone embryonic cuticle phenotype...A wing specific F 1 genetic screen was carried out using the powerful Drosophila genetic system, combined with yeast FRT/FLP and GAL4/UAS system. Form the wing phenotypes and germline clone embryonic cuticle phenotypes observed in these mutant alleles, a number of mutant alleles of known or unknown genes were isolated. Among them, fifteen mutant alleles related to Wingless signal transduction were further isolated; the arm of these mutations located were determined, and their location in the chromosome were roughly mapped.展开更多
A Flp/FRT EMS mutagenesis screen was conducted in the eye of Drosophila melanogaster on chromosome 2R to identify negative regulators of cell growth and cell division. In addition to the EMS mutation in the mosaic eye...A Flp/FRT EMS mutagenesis screen was conducted in the eye of Drosophila melanogaster on chromosome 2R to identify negative regulators of cell growth and cell division. In addition to the EMS mutation in the mosaic eye, an ark loss of function allele (ark<sup>82</sup>) was utilized to block apoptosis in the homozygous mutant cells, setting up a screen for conditional regulators of cell growth and cell division. In the present study, we focus on the characterization and mapping of one mutant that resulted from this screen, Cruella (cru). A cross between flies with the flippase enzyme directed to the developing eye and flies with the mutations cru, ark<sup>82</sup>, revealed an unusual phenotype that resulted in the homozygous mutant tissue appearing black, in contrast to the expected red. To map the location of this mutation, complementation tests against the Bloomington deficiency kit were conducted. Cru failed to complement previously characterized alleles of capping protein α (cpa). Thus, cpa<sup>cru</sup> is a novel allele of cpa and displays phenotypes similar to previously characterized alleles such as cpa 107E, cpa 69E, and cpa<sup>scrd</sup> . The human homolog, Cap Z, is conserved in humans and serves a similar role in act in filament regulation.展开更多
文摘Objective:This randomized controlled trial aimed to assess the impact of a targeted educational intervention on the awareness and practice of genetic screening and counseling among young adults in Calabar Municipality,Cross River State.Materials and Methods:Participants(aged 18-45)were randomly assigned to either an intervention group receiving structured health education or a control group receiving general health information.Stratified randomization was used between the groups.A sample size of 340 participants was recruited to detect a 20%difference in outcomes with 80%power.Data were collected using prevalidated questionnaires at baseline,immediately after the intervention,and at a 3-month follow-up.The intervention consisted of three weekly 90-min educational sessions covering genetics,the benefits of screening,and practical guidance on accessing genetic services.The primary outcomes were changes in awareness and practices related to genetic screening,with secondary outcomes focusing on attitudes and intentions toward genetic counseling.Results:Findings revealed that awareness of genetic screening was higher in the intervention group,with 65.9%of participants aware of early detection,compared to 59.4%in the control group,although the difference was not statistically significant(P=0.23).In terms of practice,42.9%of the intervention group and 40.0%of the control group engaged in genetic screening programs,with no significant difference(P=0.57).Logistic regression analysis highlighted that age,educational level,and knowledge of teratogens were significant predictors of genetic screening awareness.Participants aged 36 years and above were 1.52 times more likely to be aware(odds ratio[OR]=1.52,P=0.003),and those with tertiary education had nearly double the likelihood of awareness(OR=1.96,P<0.001).Conclusion:The study underscores the importance of targeted education in improving genetic screening awareness.
基金European Union-Next Generation EU,Through the National Recovery and Resilience Plan of the Republic of Bulgaria Project,No.BG-RRP-2.004-0008.
文摘Liver cancer,primarily hepatocellular carcinoma,remains a global health challenge with rising incidence and limited therapeutic options.Genetic factors play a pivotal role in the development and progression of liver cancer.This state-of-the-art paper provides a comprehensive review of the current landscape of genetic screening strategies for liver cancer.We discuss the genetic underpinnings of liver cancer,emphasizing the critical role of risk-associated genetic variants,somatic mutations,and epigenetic alterations.We also explore the intricate interplay between environmental factors and genetics,highlighting how genetic screening can aid in risk stratification and early detection via using liquid biopsy,and advancements in high-throughput sequencing technologies.By synthesizing the latest research findings,we aim to provide a comprehensive overview of the state-of-the-art genetic screening methods for liver cancer,shedding light on their potential to revolutionize early detection,risk assessment,and targeted therapies in the fight against this devastating disease.
文摘Objective:To detect common chromosomal aneuploidy variations in embryos from couples undergoing assisted reproductive technology and preimplantation genetic screening and their possible associations with embryo quality.Methods:In this study,359 embryos from 62 couples were screened for chromosomes 13,21,18,X,and Y by fluorescence insitu hybridization.For biopsy of blastomere,a laser was used to remove a significantly smaller portion of the zona pellucida.One blastomere was gently biopsied by an aspiration pipette through the hole.After biopsy,the embryo was immediately returned to the embryo scope until transfer.Embryo integrity and blastocyst formation were assessed on day 5.Results:Totally,282 embryos from 62 couples were evaluated.The chromosomes were normal in 199(70.57%)embryos and abnormal in 83(29.43%)embryos.There was no significant association between the quality of embryos and numerical chromosomal abnormality(P=0.67).Conclusions:Embryo quality is not significantly correlated with its genetic status.Hence,the quality of embryos determined by morphological parameters is not an appropriate method for choosing embryos without these abnormalities.
基金Supported by the National Natural Science Foundation of China,No.81901476.
文摘BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens.AIM To derive human haploid neural stem cells(haNSCs)to carry out lineage-specific screens.METHODS Human haNSCs were differentiated from human extended haESCs with the help of Y27632(ROCK signaling pathway inhibitor)and a series of cytokines to reduce diploidization.Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency.Global gene expression analysis was conducted to compare haNSCs with diploid NSCs and haESCs.Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs.Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools.RESULTS Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies,a higher efficiency in neural differentiation,a higher cell survival ratio and higher stability in haploidy maintenance.These characteristics effectively facilitated the derivation of human haNSCs.These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro.After PiggyBac transfection,there were multiple insertion sites in the human haNSCs’genome,and the insertion sites were evenly spread across all chromosomes.In addition,after the cells were treated with manganese,we were able to generate a list of manganese-induced toxicity genes,demonstrating their utility as genetic screening tools.CONCLUSION This is the first report of a generated human haploid somatic cell line with a complete genome,proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.
文摘Leukemia, like many other cancers, is thought to arise from a small population of stem cells that have the capacity to self-renewal extensively and to initiate, sustain or regenerate the disease. Elimination of the leukemia stem cells （LSCs） will likely be essential, and probably sufficient, for curing this disease.Recent studies have shown that LSCs can be derived from early hematopoietic progenitors as well as more differentiated derivatives; the key feature being these cells have acquired an increased proliferative capacity and the ability to self-renew extensively. Genes that make this possible are attractive drug targets for treating leukemia. In our laboratory we have developed a novel in vitro genetic screen that uses retroviral insertional mutagenesis as a tool for identifying genes that are able to convert both normal hematoooietic progenitors and committed mveloid progenitor cells into cells that resemble LSCs.
文摘Objectives The objective of this study was to evaluate the role of genetic screening in a Chinese woman of childbearing age with hypertrophic cardiomyopathy.Methods and Results One 39 year-old woman with presyncope for 10 years was admitted.Both of her father and her son died of sudden death and she strongly desire to get another baby. A series of clinical and laboratory studies were performed. Hypertrophic cardiomyopathy was diagnosed finally and implantable Cardioverter Defibrillator was implanted to prevent sudden cardiac death for her.Genomic DNA was isolated and the most common causal genes for hypertrophic cardiomyopathy were screened.A known pathogenetic heterozygous mutation c.700 g】a(p.Argl86Gln) in TNNI3 was found,which was not found in 100 normal control individuals matched for age,sex and geographical region.Because 50%probability of the pathogenetic mutation is inherited to the offsprings,she will get a healthy baby in vitro fertilization which the egg comes from a healthy female donor to prevent from the inherited HCM.Conclusions We found a pathogenetic TNNI3 mutation in a Chinese woman of childbearing age with hypertrophic cardiomyopathy.The genetic screening definite DNA-based diagnosis and help to design a healthy fertilization in vitro for female with hypertrophic cardiomyopathy.
基金supported by the Nanjing University Institute of Modern Biology Startup Fundthe National Natural Science Fund for Excellent Young Scientists Fund Program (Overseas)+1 种基金Fundamental Research Funds for the Central Universities (2024300391)the Jiangsu Specially Appointed Professor Fund。
文摘CRISPR technologies have revolutionized research areas ranging from fundamental science to translational medicine.CRISPR-based genetic screens offer a powerful platform for unbiased screening in various fields,such as cancer immunology.Immune checkpoint blockade(ICB)therapy has been shown to strongly affect cancer treatment.However,the currently available ICBs are limited and do not work in all cancer patients.Pooled CRISPR screens enable the identification of previously unknown immune regulators that can regulate T-cell activation,cytotoxicity,persistence,infiltration into tumors,cytokine secretion,memory formation,T-cell metabolism,and CD4^(+)T-cell differentiation.These novel targets can be developed as new immunotherapies or used with the current ICBs as new combination therapies that may yield synergistic efficacy.Here,we review the progress made in the development of CRISPR technologies,particularly technological advances in CRISPR screens and their application in novel target identification for immunotherapy.
文摘Objective To screen and identify genetic loci affecting the active zone formation in C. elegans. Methods A SYD-2：：GFP reporter was constructed and used as an active zone marker for forward genetic screen to identify genetic loci affecting the active zone formation. Results Eight isolated mutant alleles were characterized from 15,000 haploid genomes. The SYD-2：：GFP phenotypes of these mutants are mainly reflected as the changes of number, morphology, distribution of puncta and the gaps appearance. Some mutants also exhibit visible behavioral or physical phenotypes, and aldicarb resistant or sensitive phenotypes. Conclusion These mutants provide the opportunity for further systematic research on the active zone formation and the neurotransmission.
文摘The Saudi genome project started in 2013 with a great hope to improve medical care and disease prevention.Among the genes are those related to nutrition and fitness that can optimize an individual’s lifestyle.Our aim was to review the knowledge and acceptance of nutrition and fitness genetic testing to enhance the quality of life among the population of Saudi Arabia.For the study an electronic questionnaire consisting of 27 questions was prepared,and it was answered by 302 respondents.The respondents’demographics showed about 50% of respondents were aged 18-25 years and about 50% of respondents were aged 26-60 years.More than 50% of respondents were interested in having a genetic test to enhance their health,while 40% were interested in having a genetic test to enhance their fitness.Less than 50% of respondents had an understanding of the effects of coffee,macronutrition and micronutrition,elements,and enzyme activity.These results represented a contribution to the discussion on the relevance of genetic testing validity and acceptance among the population of Saudi Arabia.The results might help in producing specific guidelines on genetic testing and genomic analysis and help in the implementation of fitness and future health plans in cooperation with Saudi genome projects.Future study will focus on population structure and genetic frequency related to specific diets or fitness.
文摘The utilisation of polygenic scoring models may enhance the clinician’s ability to risk stratify an inflammatory bowel disease patient’s individual risk for venous thromboembolism(VTE)and guide the appropriate usage of VTE thromboprophylaxis,yet there is a need to validate such models in ethnically diverse populations.
文摘The advancement of Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)gene editing technology has revolutionized the comprehension of human genome,propelling molecular and cellular biology research into unexplored realms and accelerating progress in life sciences and medicine.CRISPR-based gene screening,recognized for its efficiency and practicality,is widely utilized across diverse biological fields.Aging is a multifaceted process governed by a myriad of genetic and epigenetic factors.Unraveling the genes regulating aging holds promise for understanding this intricate phenomenon and devising strategies for its assessment and intervention.This review provides a comprehensive overview of the progress in CRISPR screening and its applications in aging research,while also offering insights into future directions.CRISPR-based genetic-manipulation tools are positioned as indispensable instruments for mitigating aging and managing age-related diseases.
文摘Charcot-Marie-Tooth （CMT） disease is the most common hereditary neuropathy, with a population prevalence of 1 in 2500. CMT disease type 1A （CMT1A）, accounting for ~70% of CMT1 cases and ~ 50% of all CMT cases, is transmitted in an autosomal dominant manner. CMT1A maps to chromo- some 17pl 1.2 and is caused, in the majority of cases, by a 1.4- Mb tandem duplication that includes the peripheral myelin protein22 （PMP22） gene （Li et al., 2013）. The disease usually presents in the first 20 years of age, causing difficulty in walking or running, distal symmetrical muscle weakness and wasting, and sensory loss （van Paassen et al., 2014）.
文摘The rapidly evolving environment of assisted reproductive technology(ART)requires consideration of how new innovations are reshaping clinical practice as much as research.In particular,there are three key areas that,while full of promise,also present significant challenges:the use of artificial intelligence(AI)in embryo selection,the impact of personalized medicine on ART success rates,and the ethical considerations of genetic screening of embryos[1].This letter is meant to provoke further discussion and highlight the need for harmonized global guidelines as these advances continue to reshape the reproductive medicine environment.
基金supported by a National Research Foundation of Korea(NRF)grant funded by the Korean government(MSIT)(No.2022R1A2C2092005)the Soonchunhyang University Research Fund。
文摘Objective:Lynch syndrome(LS)increases the risk of various cancers,including colorectal cancer,endometrial cancer and gastric cancer(GC).The incidence of LS among microsatellite instability-high(MSI-H)GC and their association in South Korea remains underexplored.This study investigates LS-associated pathogenic germline variants in MSI-H GC patients using whole-exome sequencing(WES)on normal tissues.Methods:This retrospective study included patients who underwent gastrectomy for GC at Soonchunhyang University Bucheon and Cheonan Hospitals from January 2011 to October 2023.Among 1,537 patients screened for MSI status,127(8.3%)were identified as MSI-H.WES was performed on normal tissues from 123 patients.Pathogenic/likely pathogenic(P/LP)variants in mismatch repair(MMR)genes were identified using in silico models and protein loss assessments in corresponding tumor tissues.Results:Of the 127 MSI-H GC cases,characteristics aligned with typical MSI-H GC.The average age was70.02 years,with 98(77.2%)located in the lower body and 81(63.8%)of the intestinal type.All five MSI markers were positive in 46.5%of cases,whereas four markers were positive in 27.6%.Of the MSI-H GCs,10 LS candidates were identified.Three patients had known P/LP variants[MLH1(c.1758dup),MSH6(c.3261dup),MSH2(c.1241T>C)].Seven patients had variants of unknown significance(VUS)in MMR genes.Six(4.9%)patients were identified as having LS or possible LS,including one patient with the MLH1(c.1153C>T)variant previously classified as VUS but now considered LS-associated.Conclusions:This large-scale screening for LS in MSI-H GC patients using retrospective samples confirmed the lower incidence of LS than those of colorectal or endometrial cancer and GC patients in Western countries,emphasizing the need for clinical consideration in managing MSI-H GC patients.
基金supported by the National Key Research and Development Program of China(2023YFC2705900)the Fundamental Research Funds for the Central Universities(63223030)+1 种基金the National Natural Science Foundation of China(82371671,82201840,and 82301879)the Natural Science Foundation of Tianjin City(22JCZDJC00480).
文摘Trophoblast stem cells(TSCs)are placental progenitors derived from the trophectoderm or the extraembryonic ectoderm.Recently,human TSCs were established via a specific culture system[1].However,the process by which human TSCs further differentiate into terminal trophoblasts that mimic placental development remains obscure and interesting.Hence,the development of a suitable celltool to robustly study the mechanism underlying the self-renewal or differentiation of human TSCs is urgently needed.Mammalian haploid cells resemble their diploid counterparts but have a single genome,which is widely used in the genetic screening of many biological processes.However,no haploid cells for an extraembryonic lineage in humans have been derived.
基金financially supported by the National Key Research and Development Program of China (2017YFD0101001)Beijing Municipal Government Science Foundation (IDHT20170513)Peking University Institute of Advanced Agricultural Sciences。
文摘The breeding of herbicide-resistant wheat varieties has helped control weeds in wheat fields economically and effectively.Imidazolinone (IMI) herbicides are popular as they have low toxicity in mammals,are effective at small doses,and exhibit broad-spectrum herbicidal action in the field.Therefore,the isolation and genetic and molecular characterization of IMI-resistant wheat mutants will enhance weed management in wheat fields.In the present study,352 IMI-resistant plants were isolated by genetic screening from a mutant pool prepared by EMS-based random mutagenesis.Cloning of the mutated genes from the IMI-resistant plants indicated that ten taals alleles had been isolated,and mutation in one of three Ta ALS homolog genes conferred IMI resistance,and such a mutation is a dominant trait.Further analysis showed that taals-d exhibited the greatest IMI resistance,whereas taals-b exhibited the weakest resistance to IMI among three homologous taals mutants.In terms of IMI resistance,the taals triple mutant was stronger than the taals double mutants,and the taals double mutants were stronger than the single mutants,indicating a dose-dependent effect of the Ta ALS mutation on IMI resistance in wheat.Biochemical analysis indicated that the mutation in Ta ALS increased the tolerance of Ta ALS to inhibition by IMI.Our work details the genetic and molecular characterization of als wheat mutants,provides a foundation for understanding IMI resistance and breeding wheat varieties with herbicide resistance,and indicates that genetic screening using a mutagenized pool is an effective and important means of breeding crops with additional desired agricultural traits.
文摘This paper offers a general review and comparative analysis of various types of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technologies. It evaluates the strengths and weaknesses of these technologies to identify the optimal approach for conducting genetic screens. Through an extensive literature review, this paper examines CRISPR nuclease, CRISPR activation (CRISPRa), and CRISPR interference (CRISPRi) screens. This study concludes that CRISPRa and CRISPRi are more advantageous due to their use of deactivated Cas9 proteins that only over-express or deactivate genes rather than irreversibly breaking genes like CRISPRn. Notably, CRISPRa is unique in its ability to over-express genes, while the other two technologies deactivate genes. Future studies may focus on inducing multiple mutations simultaneously—both gain-of-function and gene knockout—to carry out a more complete screen that can test the combinatorial effect of genes. Likewise, targeting both exons and introns can offer a more thorough understanding of a specific phenotype.
基金Project (No.30230070) supported partially by the National Natural Science Foundation of China
文摘A wing specific F 1 genetic screen was carried out using the powerful Drosophila genetic system, combined with yeast FRT/FLP and GAL4/UAS system. Form the wing phenotypes and germline clone embryonic cuticle phenotypes observed in these mutant alleles, a number of mutant alleles of known or unknown genes were isolated. Among them, fifteen mutant alleles related to Wingless signal transduction were further isolated; the arm of these mutations located were determined, and their location in the chromosome were roughly mapped.
文摘A Flp/FRT EMS mutagenesis screen was conducted in the eye of Drosophila melanogaster on chromosome 2R to identify negative regulators of cell growth and cell division. In addition to the EMS mutation in the mosaic eye, an ark loss of function allele (ark<sup>82</sup>) was utilized to block apoptosis in the homozygous mutant cells, setting up a screen for conditional regulators of cell growth and cell division. In the present study, we focus on the characterization and mapping of one mutant that resulted from this screen, Cruella (cru). A cross between flies with the flippase enzyme directed to the developing eye and flies with the mutations cru, ark<sup>82</sup>, revealed an unusual phenotype that resulted in the homozygous mutant tissue appearing black, in contrast to the expected red. To map the location of this mutation, complementation tests against the Bloomington deficiency kit were conducted. Cru failed to complement previously characterized alleles of capping protein α (cpa). Thus, cpa<sup>cru</sup> is a novel allele of cpa and displays phenotypes similar to previously characterized alleles such as cpa 107E, cpa 69E, and cpa<sup>scrd</sup> . The human homolog, Cap Z, is conserved in humans and serves a similar role in act in filament regulation.
