Posttranscriptional regulations of different types of RNA,including rRNA,tRNA,mRNA and ncRNA are widely involved in normal physiology and diseases.m RNA,as the intermediary product between gene and protein,whose postt...Posttranscriptional regulations of different types of RNA,including rRNA,tRNA,mRNA and ncRNA are widely involved in normal physiology and diseases.m RNA,as the intermediary product between gene and protein,whose posttranscriptional regulations such as alternative splicing,alternative polyadenylation and modifications impact its coded protein expression and functions.However,the functional significance and therapeutic potential of RNA posttranscriptional regulations are not well studied due to the lack of suitable RNA engineering platforms.The discovery of a novel CRISPR-Cas system termed CRISPR-Cas13 in 2015 that specifically targets RNA templates brought a new role to CRISPR to target and edit RNA with high specificity,which opened a new era of RNA manipulations to some degree.This review will summarize the emerging applications of the catalytically inactive CRISPR-Cas13 system(CRISPR-dCas13)in mRNA engineering and highlight the prospection of the CRISPR-dCas13 system for other RNA modification regulations and its therapeutic potential.展开更多
食源性病原微生物是涉及食品安全的重要因素,传统检测方法中存在诸多局限性问题,如预处理复杂、周转时间长、灵敏度低、依赖大型仪器设备等。新发现的CRISPR (clustered regularly interspaced short palindromic repeats)技术在现阶段...食源性病原微生物是涉及食品安全的重要因素,传统检测方法中存在诸多局限性问题,如预处理复杂、周转时间长、灵敏度低、依赖大型仪器设备等。新发现的CRISPR (clustered regularly interspaced short palindromic repeats)技术在现阶段微生物检测领域出现许多新的研究进展。利用现代生物学方法基于CRISPR-Cas生物传感系统的开发,可以解决传统检测方式中的诸多问题。文章综述了依托三类Cas蛋白(Cas9、Cas12、Cas13)构建的生物传感器,并将这些生物传感器应用于食源性病原微生物的检测。这些基于CRISPR-Cas系统的传感技术有效克服了传统检测方法存在的限制,具有特异性强、灵敏度高、检测成本低的特点。文章还概述了该技术在目前研究和应用阶段遇到的问题,并对CRISPR-Cas生物传感器未来的发展方向进行了前瞻,同时提出了新的观点和可能的应用,以进一步探寻其在微生物检测领域的未来潜力。随着CRISPR-Cas系统的发展与完善,其必将在食源性微生物检测方面得到越发广泛的应用。Foodborne pathogenic microorganisms are important factors related to food safety, and traditional detection methods have many limitations. The newly discovered clustered regularly interspaced short palindromic repeats (CRISPR) technology has made many new advances in microbial detection. The use of modern biological methods based on the development of CRISPR-Cas biosensor system can provide new ideas for traditional detection methods, and solve the problems of traditional detection methods, such as complex pretreatment and long turnaround time. This article mainly reviews studies on the detection of foodborne pathogenic microorganisms based on the biosensing system with three Cas proteins (Cas9, Cas12, Cas13), which has many advantages, such as high sensitivity, high specificity, low cost and so on, breaking the limitations faced by traditional foodborne pathogenic microorganisms detection. The challenges in the current development and application of this method are summarized, and the future development prospect of the new biosensor CRISPR-Cas system is prospected and new thinking is provided for future applications to explore its potential application in microbial detection. As the CRISPR-Cas system continues to evolve and enhance, its application in identifying foodborne microbes is expected to expand significantly.展开更多
Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestat...Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.展开更多
The ongoing global pandemic of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in over 570 million infections and 6 million deaths worldwid...The ongoing global pandemic of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in over 570 million infections and 6 million deaths worldwide. Early detection and quarantine are essential to arrest the spread of the highly contagious COVID-19. High-risk groups, such as older adults and individuals with comorbidities, can present severe symptoms, including pyrexia, pertussis, and acute respiratory distress syndrome, on SARS-CoV-2 infection that can prove fatal, demonstrating a clear need for high-throughput and sensitive platforms to detect and eliminate SARS-CoV-2. CRISPR-Cas13, an emerging CRISPR system targeting RNA with high specificity and efficiency, has recently drawn much attention for COVID-19 diagnosis and treatment. Here, we summarized the current research progress on CRISPR-Cas13 in COVID-19 diagnosis and treatment and highlight the challenges and future research directions of CRISPR-Cas13 for effectively counteracting COVID-19.展开更多
文摘Posttranscriptional regulations of different types of RNA,including rRNA,tRNA,mRNA and ncRNA are widely involved in normal physiology and diseases.m RNA,as the intermediary product between gene and protein,whose posttranscriptional regulations such as alternative splicing,alternative polyadenylation and modifications impact its coded protein expression and functions.However,the functional significance and therapeutic potential of RNA posttranscriptional regulations are not well studied due to the lack of suitable RNA engineering platforms.The discovery of a novel CRISPR-Cas system termed CRISPR-Cas13 in 2015 that specifically targets RNA templates brought a new role to CRISPR to target and edit RNA with high specificity,which opened a new era of RNA manipulations to some degree.This review will summarize the emerging applications of the catalytically inactive CRISPR-Cas13 system(CRISPR-dCas13)in mRNA engineering and highlight the prospection of the CRISPR-dCas13 system for other RNA modification regulations and its therapeutic potential.
文摘食源性病原微生物是涉及食品安全的重要因素,传统检测方法中存在诸多局限性问题,如预处理复杂、周转时间长、灵敏度低、依赖大型仪器设备等。新发现的CRISPR (clustered regularly interspaced short palindromic repeats)技术在现阶段微生物检测领域出现许多新的研究进展。利用现代生物学方法基于CRISPR-Cas生物传感系统的开发,可以解决传统检测方式中的诸多问题。文章综述了依托三类Cas蛋白(Cas9、Cas12、Cas13)构建的生物传感器,并将这些生物传感器应用于食源性病原微生物的检测。这些基于CRISPR-Cas系统的传感技术有效克服了传统检测方法存在的限制,具有特异性强、灵敏度高、检测成本低的特点。文章还概述了该技术在目前研究和应用阶段遇到的问题,并对CRISPR-Cas生物传感器未来的发展方向进行了前瞻,同时提出了新的观点和可能的应用,以进一步探寻其在微生物检测领域的未来潜力。随着CRISPR-Cas系统的发展与完善,其必将在食源性微生物检测方面得到越发广泛的应用。Foodborne pathogenic microorganisms are important factors related to food safety, and traditional detection methods have many limitations. The newly discovered clustered regularly interspaced short palindromic repeats (CRISPR) technology has made many new advances in microbial detection. The use of modern biological methods based on the development of CRISPR-Cas biosensor system can provide new ideas for traditional detection methods, and solve the problems of traditional detection methods, such as complex pretreatment and long turnaround time. This article mainly reviews studies on the detection of foodborne pathogenic microorganisms based on the biosensing system with three Cas proteins (Cas9, Cas12, Cas13), which has many advantages, such as high sensitivity, high specificity, low cost and so on, breaking the limitations faced by traditional foodborne pathogenic microorganisms detection. The challenges in the current development and application of this method are summarized, and the future development prospect of the new biosensor CRISPR-Cas system is prospected and new thinking is provided for future applications to explore its potential application in microbial detection. As the CRISPR-Cas system continues to evolve and enhance, its application in identifying foodborne microbes is expected to expand significantly.
基金financially supported by the National Research Foundation of Korea(NRF-2021R1A5A1033157 for SRC program:382 Comparative medicine Disease Research Center,NRF-2021R1F1A105195313)the Research Institute of Veterinary Science,the BK21 Four for Future Veterinary Medicine Leading Education and Research Center,and a Seoul National University(SNU)grant(#550e2020005)。
文摘Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.
基金supported by Top-notch personnel from the Shanghai University of Traditional Chinese Medicine and the National Natural Science Foundation of China(No.82202922).
文摘The ongoing global pandemic of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in over 570 million infections and 6 million deaths worldwide. Early detection and quarantine are essential to arrest the spread of the highly contagious COVID-19. High-risk groups, such as older adults and individuals with comorbidities, can present severe symptoms, including pyrexia, pertussis, and acute respiratory distress syndrome, on SARS-CoV-2 infection that can prove fatal, demonstrating a clear need for high-throughput and sensitive platforms to detect and eliminate SARS-CoV-2. CRISPR-Cas13, an emerging CRISPR system targeting RNA with high specificity and efficiency, has recently drawn much attention for COVID-19 diagnosis and treatment. Here, we summarized the current research progress on CRISPR-Cas13 in COVID-19 diagnosis and treatment and highlight the challenges and future research directions of CRISPR-Cas13 for effectively counteracting COVID-19.
