单碱基编辑技术是以CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats)系统为基础衍生出的能够对基因组进行精确定点编辑的一项新技术。与CRISPR/Cas9技术系统不同,单碱基编辑系统通过在双链造成单切口后对碱基...单碱基编辑技术是以CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats)系统为基础衍生出的能够对基因组进行精确定点编辑的一项新技术。与CRISPR/Cas9技术系统不同,单碱基编辑系统通过在双链造成单切口后对碱基进行单碱基的替换,有效解决CRISPR/Cas9基因编辑系统双链断裂易产生片段插入或者缺失的缺陷,提高单碱基编辑的精确性及效率。本文介绍了胞嘧啶碱基编辑器(Cytosine Base Editor)、腺嘌呤碱基编辑器(Adenine Base Editor)以及最新的引导编辑系统(Prime Editing)的建立过程、原理、优缺点及其在动物育种中的应用,并对单碱基编辑技术发展进行展望。展开更多
碱基编辑技术是以CRISPR/Cas系统为基础开发的一种能够对基因组进行定点精准编辑的新技术,包括胞嘧啶碱基编辑系统(cytosine base editor,CBE),腺嘌呤碱基编辑系统(adenine base editor,ABE)以及引导编辑系统(primeediting,PE)。胞嘧啶...碱基编辑技术是以CRISPR/Cas系统为基础开发的一种能够对基因组进行定点精准编辑的新技术,包括胞嘧啶碱基编辑系统(cytosine base editor,CBE),腺嘌呤碱基编辑系统(adenine base editor,ABE)以及引导编辑系统(primeediting,PE)。胞嘧啶碱基编辑系统可以将基因组靶位点处的C/G转换为T/A,腺嘌呤碱基编辑系统可以将靶位点处的A/T转变为G/C,而引导编辑系统则可以实现所有12种类型(C-T、G-A、A-G、T-C、C-A、C-G、G-C、G-T、A-C、A-T、T-A、T-G)碱基的任意替换以及碱基的插入和删除。本文中系统介绍了这3种碱基编辑系统的原理、开发过程、各自的优缺点以及在作物遗传改良中的应用和发展,并展望了碱基编辑技术在农作物育种中的应用前景。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
近年来,基于成簇的规律间隔短回文重复序列及其相关系统(Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein,CRISPR/Cas)的基因编辑技术飞速发展,该系统可以利用同源定向重组(Homology directed re...近年来,基于成簇的规律间隔短回文重复序列及其相关系统(Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein,CRISPR/Cas)的基因编辑技术飞速发展,该系统可以利用同源定向重组(Homology directed repair,HDR)来完成其介导的精准编辑,但效率极低,限制了其在农业和生物医学等领域上的推广应用。基于CRISPR/Cas系统的DNA碱基编辑技术作为一种新兴的基因组编辑技术,能在不产生双链断裂的情况下实现碱基的定向突变,相对于CRISPR/Cas介导的HDR编辑具有更高的编辑效率和特异性。目前,已开发出了可将C碱基突变为T碱基的胞嘧啶碱基编辑器(Cytidine base editors,CBE),将A碱基突变为G碱基的腺嘌呤碱基编辑器(Adenine base editors,ABE),以及可实现碱基任意变换和小片段精准插入和缺失的Prime编辑器(Prime editors,PE)。另外,能实现C到G颠换的糖基化酶碱基编辑器(Glycosylase base editors,GBE)以及能同时编辑A和C两种底物的双碱基编辑器也已被开发出来。文中主要综述了几种DNA碱基编辑器的开发历程、研究进展及各自优点和局限性;介绍了DNA碱基编辑技术在生物医学以及农业中的成功应用案例,以期为DNA碱基编辑器的进一步优化和选择应用提供借鉴。展开更多
CRISPR(Clustered regularly interspaced short palindromic repeats,规律间隔的成簇短回文重复序列)/Cas9(CRISPR-associated protein 9,CRISPR相关蛋白9)系统是一种强大的基因编辑工具,因其成本低、可操作性强、编辑效率高等优点,已...CRISPR(Clustered regularly interspaced short palindromic repeats,规律间隔的成簇短回文重复序列)/Cas9(CRISPR-associated protein 9,CRISPR相关蛋白9)系统是一种强大的基因编辑工具,因其成本低、可操作性强、编辑效率高等优点,已被大量应用于农作物改良中。文章综述了CRISPR基因编辑系统的发现、分类、作用机制以及CRISPR/Cas9在农作物改良上的应用,并展望其未来的发展前景。展开更多
Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop breeding.Four new types of prime editors(PEs)named PE6a–d were recently generated using evolved and engineered reverse transcrip...Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop breeding.Four new types of prime editors(PEs)named PE6a–d were recently generated using evolved and engineered reverse transcriptase(RT)variants from three different sources.In this study,we tested the editing efficiencies of four PE6 variants and two additional PE6 constructs with double-RT modules in transgenic rice(Oryza sativa)plants.PE6c,with an evolved and engineered RT variant from the yeast Tf1 retrotransposon,yielded the highest prime-editing efficiency.The average fold change in the editing efficiency of PE6c compared with PEmax exceeded 3.5 across 18 agronomically important target sites from 15 genes.We also demonstrated the feasibility of using two RT modules to improve prime-editing efficiency.Our results suggest that PE6c or its derivatives would be an excellent choice for prime editing in monocot plants.In addition,our findings have laid a foundation for prime-editing-based breeding of rice varieties with enhanced agronomically important traits.展开更多
文摘单碱基编辑技术是以CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats)系统为基础衍生出的能够对基因组进行精确定点编辑的一项新技术。与CRISPR/Cas9技术系统不同,单碱基编辑系统通过在双链造成单切口后对碱基进行单碱基的替换,有效解决CRISPR/Cas9基因编辑系统双链断裂易产生片段插入或者缺失的缺陷,提高单碱基编辑的精确性及效率。本文介绍了胞嘧啶碱基编辑器(Cytosine Base Editor)、腺嘌呤碱基编辑器(Adenine Base Editor)以及最新的引导编辑系统(Prime Editing)的建立过程、原理、优缺点及其在动物育种中的应用,并对单碱基编辑技术发展进行展望。
文摘碱基编辑技术是以CRISPR/Cas系统为基础开发的一种能够对基因组进行定点精准编辑的新技术,包括胞嘧啶碱基编辑系统(cytosine base editor,CBE),腺嘌呤碱基编辑系统(adenine base editor,ABE)以及引导编辑系统(primeediting,PE)。胞嘧啶碱基编辑系统可以将基因组靶位点处的C/G转换为T/A,腺嘌呤碱基编辑系统可以将靶位点处的A/T转变为G/C,而引导编辑系统则可以实现所有12种类型(C-T、G-A、A-G、T-C、C-A、C-G、G-C、G-T、A-C、A-T、T-A、T-G)碱基的任意替换以及碱基的插入和删除。本文中系统介绍了这3种碱基编辑系统的原理、开发过程、各自的优缺点以及在作物遗传改良中的应用和发展,并展望了碱基编辑技术在农作物育种中的应用前景。
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.
文摘近年来,基于成簇的规律间隔短回文重复序列及其相关系统(Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein,CRISPR/Cas)的基因编辑技术飞速发展,该系统可以利用同源定向重组(Homology directed repair,HDR)来完成其介导的精准编辑,但效率极低,限制了其在农业和生物医学等领域上的推广应用。基于CRISPR/Cas系统的DNA碱基编辑技术作为一种新兴的基因组编辑技术,能在不产生双链断裂的情况下实现碱基的定向突变,相对于CRISPR/Cas介导的HDR编辑具有更高的编辑效率和特异性。目前,已开发出了可将C碱基突变为T碱基的胞嘧啶碱基编辑器(Cytidine base editors,CBE),将A碱基突变为G碱基的腺嘌呤碱基编辑器(Adenine base editors,ABE),以及可实现碱基任意变换和小片段精准插入和缺失的Prime编辑器(Prime editors,PE)。另外,能实现C到G颠换的糖基化酶碱基编辑器(Glycosylase base editors,GBE)以及能同时编辑A和C两种底物的双碱基编辑器也已被开发出来。文中主要综述了几种DNA碱基编辑器的开发历程、研究进展及各自优点和局限性;介绍了DNA碱基编辑技术在生物医学以及农业中的成功应用案例,以期为DNA碱基编辑器的进一步优化和选择应用提供借鉴。
文摘CRISPR(Clustered regularly interspaced short palindromic repeats,规律间隔的成簇短回文重复序列)/Cas9(CRISPR-associated protein 9,CRISPR相关蛋白9)系统是一种强大的基因编辑工具,因其成本低、可操作性强、编辑效率高等优点,已被大量应用于农作物改良中。文章综述了CRISPR基因编辑系统的发现、分类、作用机制以及CRISPR/Cas9在农作物改良上的应用,并展望其未来的发展前景。
基金supported by grants from the National Key Research and Development Program of China (2023YFD1202905)the National Natural Science Foundation of China (U19A2022)
文摘Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop breeding.Four new types of prime editors(PEs)named PE6a–d were recently generated using evolved and engineered reverse transcriptase(RT)variants from three different sources.In this study,we tested the editing efficiencies of four PE6 variants and two additional PE6 constructs with double-RT modules in transgenic rice(Oryza sativa)plants.PE6c,with an evolved and engineered RT variant from the yeast Tf1 retrotransposon,yielded the highest prime-editing efficiency.The average fold change in the editing efficiency of PE6c compared with PEmax exceeded 3.5 across 18 agronomically important target sites from 15 genes.We also demonstrated the feasibility of using two RT modules to improve prime-editing efficiency.Our results suggest that PE6c or its derivatives would be an excellent choice for prime editing in monocot plants.In addition,our findings have laid a foundation for prime-editing-based breeding of rice varieties with enhanced agronomically important traits.
