Development of efficient targeted insertion mediated by CRISPR-Cas12a and homology-directed repair in maize.

ABSTRACT: Targeted insertion (TIN) of transgenic trait cassettes has the potential to reduce timeline and cost for GM product development and commercialization. Precise genome engineering has made remarkable progress over the last several decades, particularly with the use of site-directed nucleases as core editing machinery. However, there are still many critical factors that can impact TIN efficiency including insertion site selection, nuclease optimization and expression, donor vector design, gene delivery, and stable event regeneration. Here, we develop workflows for target site sequence identification and gRNA screening for CRISPR-Cas12a system and demonstrate its successful application for TIN in maize with donor sequences up to 10 kilobase pairs (kb) in size. We first prioritize genomic regio

SUBMITTER: Barco B 

PROVIDER: S-EPMC12711819 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature