本研究首次在热带假丝酵母中开发了tRNA-gRNA策略,并基于该策略构建了CRISPR-dCas9基因调控系统.论文主要结果如下:(1)内源tRNA编码基因的挖掘.利用生物信息学工具tRNAscan-SE对热带假丝酵母ATCC 20336基因组序列进行分析,获得全基因组序列中tRNA的编码序列.选择一条长度为71 bp内源tRNAGly序列,与已报道的异源tRNAGly...
Here, we report a gRNA-tRNA array for CRISPR-Cas9 (GTR-CRISPR) for multiplexed engineering of. Using reported gRNAs shown to be effective, this system enables simultaneous disruption of 8 genes with 87% efficiency. We further report an accelerated Lightning GTR-CRISPR that avoids the cloning ...
由于参与翻译DNA片段的tRNA的基本识别单位是3个碱基,因此如果被剪目标处,在修复过程中出现以3个碱基倍数的增减,则意味着原DNA翻译成蛋白质的过程仍将继续,只不过是相比原来增减对应数量的氨基酸。下列表述中,符合文意的一项是: A.DNA在修复过程中出现以3个碱基倍数的增减,会导致“翻译过程”被迫中断 B.通过gRNA与...
目的:探究巨细胞病毒(CMV)启动子驱动转运RNA-导向RNA(tRNA-gRNA)的表达是否能够在青鳉细胞系及胚胎中产生功能性的gRNA并诱导基因的突变。 创新点:利用CMV启动子驱动tRNA-gRNA的表达在青鳉细胞系及胚胎中产生功能性的gRNA,结合Cas9系统诱导了青鳉胚胎和细胞系中的基因突变,为在鱼类胚胎和细胞系中进行基因编辑提供了一...
Poly-tRNA-gRNAEmbryosFish cellsGeneration of mutants with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is commonly carried out in fish species by co-injecting a mixture ofCas9messenger RNA (mRNA) or protein and transcribed guide RNA (gRNA). ...
tRNA-gRNA arraytriplex sequenceSimultaneously, multiplexed genome engineering and targeting multiple genomic loci are valuable to elucidating gene interactions and characterizing genetic networks that affect phenotypes. Here, we developed a general CRISPR-based platform to perform four functions ...
To develop an easy and robust method for creating genetically stable and easily detectable Arabidopsis mutants, we adopted the polycistronic tRNA-gRNA CRISPR/Cas9 (PTG/Cas9) system, a multiplex gene-editing tool in rice, with PTOX as the reporter gene. The PTG/Cas9 system has a great ...
Our results demonstrate that tRNA promoter-mediated gRNA expressions are reliable and efficient in genome editing in A. niger.Letian SongJean-Paul OuedraogoMagdalena KolbuszThi Truc Minh NguyenAdrian TsangPLoS ONE (v.1;2006)Song L, Ouedraogo JP, Kolbusz M, Nguyen TTM, Tsang A. 2018. Efficient ...
gRNAtRNA arraySCNTMultiplex gene modifications are highly required for various fields of porcine research. In many species, the CRISPR/Cas9 system has been widely applied for genomic editing and provides a potential tool for introducing multiplex genome mutations simultaneously. Here, we present a ...
Polycistronic tRNA–gRNA CRISPR/Cas9 systemPTOX (plastid terminal oxidase)Cas9-free deletion mutantTo develop an easy and robust method for creating genetically stable and easily detectable Arabidopsismutants, we adopted the polycistronic tRNA–gRNA CRISPR/Cas9 (PTG/Cas9) system, a multiplex......