| Literature DB >> 30610628 |
Kabin Xie1, Yinong Yang2.
Abstract
The CRISPR-Cas9 system has become a powerful and popular tool for genome editing due to its efficiency and simplicity. Multiplex genome editing is an important feature of the CRISPR-Cas9 system and requires simultaneous expression of multiple guide RNAs (gRNAs). Here we describe a general method to efficiently produce many gRNAs from a single gene transcript based on the endogenous tRNA-processing system. A step-by-step protocol is provided for the design and construction of the polycistronic tRNA-gRNA (PTG) gene. The PTG method has been demonstrated to be highly efficient for multiplex genome editing in various plant, animal, and microbial species.Entities:
Keywords: CRISPR-Cas9; Genome editing; Multiplex; gRNA; tRNA
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Year: 2019 PMID： 30610628 DOI： 10.1007/978-1-4939-8991-1_5
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745