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Engineering Introns to Express RNA Guides for Cas9- and Cpf1-Mediated Multiplex Genome Editing.

Mol Plant. 2018-04; 
DingDan,ChenKaiyuan,ChenYuedan,LiHong,XieK
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Gene Synthesis … pRGEB3345. The Lachnospiraceae Cpf1 (LbCpf1) was obtained from Addgene (Plasmid #69988) (Zetsche et al., 2015), and the rice codon optimized Francisella novidica Cpf1 (FnCpf1) was synthesized at GenScript. Both LbCpf1 … Get A Quote

摘要

The clustered regularly interspaced short palindromic repeat (CRISPR) system has emerged as the revolutionary platform for DNA targeting. This system uses a site-specific RNA guide to direct a CRISPR effector (e.g., Cas9 and Cpf1) to a DNA target. Here, we elaborate a general strategy to simultaneously express multiple guide RNAs (gRNA) and CRISPR RNAs (crRNA) from introns of Cas9 and Cpf1. This method utilizes the endogenous tRNA processing system or crRNA processing activity of Cpf1 to cleave the spliced intron that contains tRNA-gRNA polycistron or crRNA-crRNA array. We demonstrated that the tRNA-gRNA intron is able to fuse with Cas9 as one gene. Such a hybrid gene could be expressed using one polymerase... More

关键词

CRISPR,Cas9,Cpf1,intron,multi