Conditional knockdown of OsMLH1 to improve plant prime editing systems without disturbing fertility in rice.

ABSTRACT:

Background

High-efficiency prime editing (PE) is desirable for precise genome manipulation. The activity of mammalian PE systems can be largely improved by inhibiting DNA mismatch repair by coexpressing a dominant-negative variant of MLH1. However, this strategy has not been widely used for PE optimization in plants, possibly because of its less conspicuous effects and inconsistent performance at different sites.

Results

We show that direct RNAi knockdown of OsMLH1 in an ePE5c system increases the efficiency of our most recently updated PE tool by 1.30- to 2.11-fold in stably transformed rice cells, resulting in as many as 85.42% homozygous mutants in the T₀ generation. The high specificity of ePE5c is revealed by whole-genome sequencing. To overcome the partial s

SUBMITTER: Liu X 

PROVIDER: S-EPMC11110357 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature