The heat is on: a simple method to increase genome editing efficiency in plants.

ABSTRACT:

Background

Precision genome mutagenesis using CRISPR/Cas has become the standard method to generate mutant plant lines. Several improvements have been made to increase mutagenesis efficiency, either through vector optimisation or the application of heat stress.

Results

Here, we present a simplified heat stress assay that can be completed in six days using commonly-available laboratory equipment. We show that three heat shocks (3xHS) efficiently increases indel efficiency of LbCas12a and Cas9, irrespective of the target sequence or the promoter used to express the nuclease. The generated indels are primarily somatic, but for three out of five targets we demonstrate that up to 25% more biallelic mutations are transmitted to the progeny when heat is applied compared to non-heat

SUBMITTER: Blomme J 

PROVIDER: S-EPMC8951696 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature