A Multiplexed CRISPR/Cas9 Editing System Based on the Endogenous tRNA Processing

Methods in Molecular Biology
Kabin Xie, Yinong Yang

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.

References

Jan 12, 2020·Journal of Applied Genetics·Luiza Chojnacka-Puchta, Dorota Sawicka

Related Concepts

System Process
Genes
CRISPR-Cas Systems
Gene Editing
Transfer RNA
Teniposide
Clustered Regularly Interspaced Short Palindromic Repeats
Species
Transcript
RNA, Guide

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