CRISPR/Cas9介导靶向敲除拟南芥BRI1突变体的鉴定

以拟南芥(Arabidopsis thaliana)油菜素内酯受体BRI1为目的基因,利用CRISPR/Cas9基因编辑技术定向编辑拟南芥BRI1,以期获得更多BRI1的突变体,为后续BRI1功能的进一步深入研究奠定基础。通过筛选转基因植株,对编辑后的BRI1进行测序分析,结果显示该突变体中BRI1基因序列由于新碱基的插入导致提前终止。同BRI1强突变体bri1-710一样,相比于野生型对照均对BL处理不敏感,但相比于bri1-710,该突变体植株较大,暗示BRI1 N端可能在BR信号途径中有重要作用。因此该研究可为后续进一步研究拟南芥及其他同源物种的BRI1功能提供可靠的参考依据。

Identification of Knockout of BRI1 Mutant in Arabidopsis Mediated by CRISPR/Cas9

CRISPR/Cas9 gene editing technology was used to specificity edit the BRI1 of Arabidopsis thaliana to creation new BRI1 mutants.We analyzed the sequence of BRI1 in the obtained transgenic plants,the results showed that as one new BRI1 mutant due to the insertion of new bases,the normal BRI1 protein is terminated prematurely.This mutant have the similar phenotype as bri1-710,and both mutants show impaired response to BL treatment compared with wild-type.Those results suggested that the N-terminal of BRI1 may play a role in the BR signaling pathway.Therefore,this study could provide a reliable reference for further studies on the BRI1 function of A.thaliana and other homologous species.