To ensure that all proteins and RNAs are expressed in one transcript, multiple gRNAs were constructed in a tandemly arrayed tRNA (transfer RNA)-gRNA architecture, and the triplex sequence was cloned between the protein-coding sequences and the tRNA-gRNA array. By leveraging this s...
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...
tRNAGly二级结构,推测出潜在核糖核酸酶P和核糖核酸酶Z切割位点.(2)RNA聚合酶Ⅲ型启动子功能鉴定及tRNA-gRNA策略的开发.在CRISPR-Cas9系统中,利用tRNAGly启动子表达靶向单基因的gRNA,乳清酸核苷-5′-磷酸脱羧酶编码基因(URA3)单基因编辑效率为100%.利用该启动子,开发了一种从单个顺反子基因表达多条gRNAs的tRNA-...
Medaka (Oryzias latipes)Gene editingPoly-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 ...
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 ...
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......
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 ...