Never-germinating Arabidopsis seeds with LbCas12a-induced mutations in 6 clade A type 2C protein phosphatase genes.

ABSTRACT: Cas12 nucleases, such as Cas12a, Cas12i, and Cas12f, are genome-editing tools that possess several unique attributes. However, the potential of various Cas12 variants for multiplex genome editing in Arabidopsis (Arabidopsis thaliana) remains insufficiently characterized. In this report, we systematically evaluated 18 additional targets and demonstrated that the LbCas12a variant carrying D156R and E795L mutations exhibits minimal target bias. Furthermore, we achieved an editing efficiency of at least 73.8% (45/61) in generating T1 homozygous sextuple mutants, with more than half of these mutants exhibiting a complete seed germination arrest phenotype. Comparative analysis of 7 LbCas12a variants revealed that the optimization of nuclear localization sequences, rather than codon usage, is fun

SUBMITTER: Xin C 

PROVIDER: S-EPMC12301882 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature