With the ever-increasing human population and deteriorating environmental conditions,there is an urgent need to breed environmentally friendly and resource-conserving rice cultivars to achieve sustainable agricultural...With the ever-increasing human population and deteriorating environmental conditions,there is an urgent need to breed environmentally friendly and resource-conserving rice cultivars to achieve sustainable agricultural development and food security.However,conventional rice improvement strategies,such as hybrid breeding,are time-consuming and laborious processes and may not be able to keep pace with increasing food demand in the future.Targeted genome-editing technologies,especially clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated protein(CRISPR/Cas),permit efficient targeted genome modification and offer great promise for the creation of desired plants with higher yield,improved grain quality,and resistance to herbicides,diseases,and insect pests.There is also great potential for tapping heterosis using the CRISPR/Cas technology.In this review,we focus on the most essential applications of CRISPR/Cas genome editing to rice genetic improvement,considering traits such as yield,quality,and herbicide,disease and insect-pest resistance.We discuss applications of CRISPR/Cas to the exploitation of heterosis.Finally,we outline perspectives for future rice breeding using genome-editing technologies.展开更多
Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most impo...Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most important strategy for managing the disease.However,studies on the CR gene functions are quite limited.In this study,we have conducted investigations into the temporal,structural,and interacting features of a newly cloned CR gene,Rcr1,using CRISPR/Cas9 technology.For temporal functionality,we developed a novel CRISPR/Cas9-based binary vector,pHHIGR-Hsp18.2,to deliver Rcr1 into a susceptible canola line(DH12075)and observed that early expression of Rcr1 is critical for conferring resistance.For structural functionality,several independent mutations in specific domains of Rcr1 resulted in loss-offunction,highlighting their importance for CR phenotype.In the study of the interacting features of Rcr1,a cysteine protease gene and its homologous allele in canola were successfully disrupted via CRISPR/Cas9 as an interacting component with Rcr1 protein,resulting in the conversion from clubroot resistant to susceptible in plants carrying intact Rcr1.These results indicated an indispensable role of these two cysteine proteases in Rcr1-mediated resistance response.This study,the first of its kind,provides valuable insights into the functionality of Rcr1.Further,the new vector p HHIGR-Hsp18.2 demonstrated an inducible feature on the removal of add-on traits,which should be useful for functional genomics and other similar research in brassica crops.展开更多
Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase i...Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase in monounsaturated fatty acids,particularly oleic acid,enhances the oxidative stability and nutritional value of edible oil(Chen et al.2021).展开更多
The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of t...The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.展开更多
The current major issue in improving detection sensitivity and selectivity is to design an electrochemical sensor that does not require PCR amplification for nucleic acid identification and measurement. Because of the...The current major issue in improving detection sensitivity and selectivity is to design an electrochemical sensor that does not require PCR amplification for nucleic acid identification and measurement. Because of their great sensitivity, precision, and simplicity of downsizing, electrochemical biosensors have emerged as a research hotspot in the field of nucleic acid detection. The CRISPR/Cas12 system has emerged as a potent tool for nucleic acid detection due to its powerful cleavage activity and selectivity. Specific electrode changes combined with the CRISPR/Cas12 system can greatly improve the performance of electrochemical biosensors. In this study, the design concepts of electrochemical biosensors based on the CRISPR/Cas12 system and their application advancements in nucleic acid detection are discussed.展开更多
Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated(Cas)systems are becoming powerful tools for disease biomarkers detection.Due to the specific recognition,cis-cleavage and nonspecific...Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated(Cas)systems are becoming powerful tools for disease biomarkers detection.Due to the specific recognition,cis-cleavage and nonspecific trans-cleavage capabilities,CRISPR/Cas systems have implemented the detection of nucleic acid targets(DNA and RNA)as well as non-nucleic acid targets(e.g.,proteins,exosomes,cells,and small molecules).In this review,we first summarize the principles and characteristics of various CRISPR/Cas systems,including CRISPR/Cas9,Cas12,Cas13 and Cas14 systems.Then,various types of applications of CRISPR/Cas systems used in detecting nucleic and non-nucleic acid targets are introduced emphatically.Finally,the prospects and challenges of their applications in biosensing are discussed.展开更多
Infectious diseases are a serious threat to human health,and accurate,rapid and convenient early detection of pathogens is the first step of active treatment.Technologies that detect pathogens have advanced significan...Infectious diseases are a serious threat to human health,and accurate,rapid and convenient early detection of pathogens is the first step of active treatment.Technologies that detect pathogens have advanced significantly because of the development of fundamental disciplines and the integration of multidisciplinary fields.Among these technologies,nucleic acid detection technology is preferred because of its rapid measurement,accuracy and high sensitivity.The CRISPR/Cas system,consisting of Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)and CRISPR‐associated(Cas),is an adaptive immune system that specifically recognizes,binds and cleaves exogenous invasive nucleic acids.The CRISPR/Cas system is widely found in bacteria and archaea.Researchers have developed nucleic acid detection technologies with single‐molecule sensitivity,single‐base precision specificity,portability and low cost based on the specific cleavage and trans‐cleavage activities of the CRISPR/Cas system.The next generation of in‐vitro diagnostics is shifting to nucleic acid technology because this technology shows promise in a wide range of applications in resource‐constrained environments.In this review,the development and mechanism of the CRISPR/Cas system are presented together with representative CRISPR/Cas applications in nucleic acid detection.Additionally,the review summarizes future perspectives and trends of the CRISPR/Cas system in nucleic acid detection.展开更多
基金supported by the National Natural Science Foundation of China(U20A2030)Central Public-interest Scientific Institution Basal Research Fund(Y2020YJ12 and Y2020XK17)+1 种基金Key Research and Development Program of China National Rice Research Institute(CNRRI-2020-01)Foreign Cooperation Project of Ningxia Academy of Agricultural and Forestry Institute(DW-X-2018004)。
文摘With the ever-increasing human population and deteriorating environmental conditions,there is an urgent need to breed environmentally friendly and resource-conserving rice cultivars to achieve sustainable agricultural development and food security.However,conventional rice improvement strategies,such as hybrid breeding,are time-consuming and laborious processes and may not be able to keep pace with increasing food demand in the future.Targeted genome-editing technologies,especially clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated protein(CRISPR/Cas),permit efficient targeted genome modification and offer great promise for the creation of desired plants with higher yield,improved grain quality,and resistance to herbicides,diseases,and insect pests.There is also great potential for tapping heterosis using the CRISPR/Cas technology.In this review,we focus on the most essential applications of CRISPR/Cas genome editing to rice genetic improvement,considering traits such as yield,quality,and herbicide,disease and insect-pest resistance.We discuss applications of CRISPR/Cas to the exploitation of heterosis.Finally,we outline perspectives for future rice breeding using genome-editing technologies.
基金supported by the Genomics Initiative of Agriculture and Agri-Food Canada。
文摘Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most important strategy for managing the disease.However,studies on the CR gene functions are quite limited.In this study,we have conducted investigations into the temporal,structural,and interacting features of a newly cloned CR gene,Rcr1,using CRISPR/Cas9 technology.For temporal functionality,we developed a novel CRISPR/Cas9-based binary vector,pHHIGR-Hsp18.2,to deliver Rcr1 into a susceptible canola line(DH12075)and observed that early expression of Rcr1 is critical for conferring resistance.For structural functionality,several independent mutations in specific domains of Rcr1 resulted in loss-offunction,highlighting their importance for CR phenotype.In the study of the interacting features of Rcr1,a cysteine protease gene and its homologous allele in canola were successfully disrupted via CRISPR/Cas9 as an interacting component with Rcr1 protein,resulting in the conversion from clubroot resistant to susceptible in plants carrying intact Rcr1.These results indicated an indispensable role of these two cysteine proteases in Rcr1-mediated resistance response.This study,the first of its kind,provides valuable insights into the functionality of Rcr1.Further,the new vector p HHIGR-Hsp18.2 demonstrated an inducible feature on the removal of add-on traits,which should be useful for functional genomics and other similar research in brassica crops.
基金supported by the Science and Technology Innovation Talents in Universities of Henan Province,China(24HASTIT053)the National Natural Science Foundation of China(32172041)+1 种基金the Natural Science Foundation of Henan Province,China(232300421026)the Science and Technology Innovation 2030,China(2022ZD0402001-04)。
文摘Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase in monounsaturated fatty acids,particularly oleic acid,enhances the oxidative stability and nutritional value of edible oil(Chen et al.2021).
基金supported by the National Natural Science Foundation of China(32001532 and 31860411)the National Key Research and Development Program of China,(2022YFF1000020)+1 种基金Hunan Seed Industry Innovation Project(2021NK1012)the Yunnan Tobacco Company Project(2020530000241009)。
文摘The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.
文摘The current major issue in improving detection sensitivity and selectivity is to design an electrochemical sensor that does not require PCR amplification for nucleic acid identification and measurement. Because of their great sensitivity, precision, and simplicity of downsizing, electrochemical biosensors have emerged as a research hotspot in the field of nucleic acid detection. The CRISPR/Cas12 system has emerged as a potent tool for nucleic acid detection due to its powerful cleavage activity and selectivity. Specific electrode changes combined with the CRISPR/Cas12 system can greatly improve the performance of electrochemical biosensors. In this study, the design concepts of electrochemical biosensors based on the CRISPR/Cas12 system and their application advancements in nucleic acid detection are discussed.
基金This work was financially supported by the National Natural Science Foundation of China(Nos.81771968 and 22204104)the Shanghai Sailing Program(No.21YF1444900)+3 种基金the Shanghai Municipal Natural Science Foundation(No.22ZR1459600)the Medical-Engineering Joint Funds from the Shanghai Jiao Tong University(Nos.YG2023ZD07 and YG2021QN23)the Foundation of Shanghai Municipal Health Commission(No.2022JC002)the Innovative Research Team of High-Level Local Universities in Shanghai,China.
文摘Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated(Cas)systems are becoming powerful tools for disease biomarkers detection.Due to the specific recognition,cis-cleavage and nonspecific trans-cleavage capabilities,CRISPR/Cas systems have implemented the detection of nucleic acid targets(DNA and RNA)as well as non-nucleic acid targets(e.g.,proteins,exosomes,cells,and small molecules).In this review,we first summarize the principles and characteristics of various CRISPR/Cas systems,including CRISPR/Cas9,Cas12,Cas13 and Cas14 systems.Then,various types of applications of CRISPR/Cas systems used in detecting nucleic and non-nucleic acid targets are introduced emphatically.Finally,the prospects and challenges of their applications in biosensing are discussed.
基金Foundation of the Innovation Academy for Green Manufacture Institute,Chinese Academy of Sciences,Grant/Award Number:IAGM2020C31National Key Research and Development Program of China,Grant/Award Numbers:2019YFC1606600,2019YFC1606602。
文摘Infectious diseases are a serious threat to human health,and accurate,rapid and convenient early detection of pathogens is the first step of active treatment.Technologies that detect pathogens have advanced significantly because of the development of fundamental disciplines and the integration of multidisciplinary fields.Among these technologies,nucleic acid detection technology is preferred because of its rapid measurement,accuracy and high sensitivity.The CRISPR/Cas system,consisting of Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)and CRISPR‐associated(Cas),is an adaptive immune system that specifically recognizes,binds and cleaves exogenous invasive nucleic acids.The CRISPR/Cas system is widely found in bacteria and archaea.Researchers have developed nucleic acid detection technologies with single‐molecule sensitivity,single‐base precision specificity,portability and low cost based on the specific cleavage and trans‐cleavage activities of the CRISPR/Cas system.The next generation of in‐vitro diagnostics is shifting to nucleic acid technology because this technology shows promise in a wide range of applications in resource‐constrained environments.In this review,the development and mechanism of the CRISPR/Cas system are presented together with representative CRISPR/Cas applications in nucleic acid detection.Additionally,the review summarizes future perspectives and trends of the CRISPR/Cas system in nucleic acid detection.
