Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 100 (4), 863-872

A CRISPR/LbCas12a-based Method for Highly Efficient Multiplex Gene Editing in Physcomitrella Patens

Affiliations

A CRISPR/LbCas12a-based Method for Highly Efficient Multiplex Gene Editing in Physcomitrella Patens

Xiaojun Pu et al. Plant J.

Abstract

Due to their high efficiency, specificity, and flexibility, programmable nucleases, such as those of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a (Cpf1) system, have greatly expanded the applicability of editing the genomes of various organisms. Genes from different gene families or genes with redundant functions in the same gene family can be examined by assembling multiple CRISPR RNAs (crRNAs) in a single vector. However, the activity and efficiency of CRISPR/Cas12a in the non-vascular plant Physcomitrella patens are largely unknown. Here, we demonstrate that LbCas12a together with its mature crRNA can target multiple loci simultaneously in P. patens with high efficiency via co-delivery of LbCas12a and a crRNA expression cassette in vivo. The mutation frequencies induced by CRISPR/LbCas12a at a single locus ranged from 26.5 to 100%, with diverse deletions being the most common type of mutation. Our method expands the repertoire of genome editing tools available for P. patens and facilitates the creation of loss-of-function mutants of multiple genes from different gene families.

Keywords: Physcomitrella patens; CRISPR/Cas12a; CRISPR/Cas9; genome editing; technical advance.

Similar articles

See all similar articles

Cited by 1 PubMed Central articles

Publication types

LinkOut - more resources

Feedback