ArticleFrontiers in plant science2024
Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy.
Article in Frontiers in plant science, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 6 citations in OpenAlex.
- SSA-mediated selection marker gene activation enhances relative gene targeting efficiency in plants.Horticulture research · 2025Article
- Exonuclease-fused CRISPR-cas system enhances targeted genome editing for functional genomics in soybean.BMC plant biology · 2025Article
- Double step screening using endogenous marker improves relative gene targeting efficiency in Arabidopsis.Scientific reports · 2024Article
- Cas12a-mediated gene targeting by sequential transformation strategy in Arabidopsis thaliana.BMC plant biology · 2024Article
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Authors and funding
7 authors at 5 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
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.
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Registered trials
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