Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy.
Front Plant Sci
; 15: 1360925, 2024.
Article
de En
| MEDLINE
| ID: mdl-38545386
ABSTRACT
Gene targeting (GT) is a promising tool for precise manipulation of genome sequences, however, GT in seed plants remains a challenging task. The simple and direct way to improve the efficiency of GT via homology-directed repair (HDR) is to increase the frequency of double-strand breaks (DSBs) at target sites in plants. Here we report an all-in-one approach of GT in Arabidopsis by combining a transcriptional and a translational enhancer for the Cas expression. We find that facilitating the expression of Cas9 and Cas12a variant by using enhancers can improve DSB and subsequent knock-in efficiency in the Arabidopsis genome. These results indicate that simply increasing Cas protein expression at specific timings - egg cells and early embryos - can improve the establishment of heritable GTs. This simple approach allows for routine genome engineering in plants.
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Langue:
En
Journal:
Front Plant Sci
Année:
2024
Type de document:
Article
Pays d'affiliation:
Chine
Pays de publication:
Suisse