Systems and methods for plant genome editing using Cas 12a orthologs

Abstract

A non-naturally occurring heterologous CRISPR-Cas12a genomic editing system is described, including or encoding at least one Cas12a ortholog endonuclease selected from among Lb5Cas12a, CMaCas12a, BsCas12a, BoCas12a, MlCas12a, Mb2Cas12a, MbCas12a TsCas12a, and MAD7® ErCas12a endonucleases. The ortholog endonucleases enable genomic editing of plants at low temperatures, e.g., below 25° C. The CRISPR-Cas12a system can be employed for targeting PAM sites such as TTN, TTV, TTTV, NTTV, TATV, TATG, TATA, YTTN, GTTA, and GTTC, utilizing corresponding gRNAs, and is readily adapted to multiplexed editing operations of plants such as rice and other food crops.

CPC Classifications