Enhancing gene editing and gene targeting efficiencies in Arabidopsis thaliana by using an intron-containing version of ttLbCas12a

编号 040037503

推送时间 20221226

研究领域 森林培育 

年份 2022 

类型 期刊 

语种 英语

标题 Enhancing gene editing and gene targeting efficiencies in Arabidopsis thaliana by using an intron-containing version of ttLbCas12a

来源期刊 Plant Biotechnology Journal

期 第375期

发表时间 20221116

关键词 CRISPR/Cas12a;  ttLBCas12a;  gene editing;  gene targeting;  intron; 

摘要 The CRISPR/Cas12a system can be used in a wide range of organisms. However, in plants, efficient editing by Cas12a was hampered due to the lower temperatures required for plant cultivation (Malzahn et al., 2019). Recently, we engineered an improved, temperature-tolerant variant of Cas12a from Lachnospiraceae bacterium, ttLbCas12a, which has shown dramatically increased editing efficiencies compared to LbCas12a in Arabidopsis at the standard cultivation temperature of 22°C (Schindele and Puchta, 2020). Furthermore, we could show that, by using ttLbCas12a, gene targeting (GT) efficiencies can be elevated considerably, both in Arabidopsis and tobacco (Huang et al., 2021;
Merker et al., 2020). Nevertheless, even when using ttLbCas12a, some target sites remain highly resistant to editing. Evidence is accumulating that especially heterochromatic sites are more
recalcitrant to editing (Weiss et al., 2022). Recently, it was demonstrated that the incorporation of 13 introns into the Cas9 coding sequence dramatically improves the editing efficiency in plants (Grützner et al., 2021). Based on these findings, an intron-containing ttLbCas12a construct (ttLbCas12a-i) was established to investigate whether editing in plants could be further improved.

服务人员 孙小满

服务院士 尹伟伦

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