Whole-genome sequencing data of plant edited by Mb2Cas12a

We established an efficient genome editing system in cotton mediated by CRISPR/Mb2Cas12a. First, we designed three vectors using various expression cassettes to verify their applicability in cotton.We have successfully obtained gossypol-free plants and leaf-bleaching plants separately, proving that all three vectors can perform efficient editing in cotton. Furthermore, we have revealed that Mb2Cas12a exhibited high precision through whole-genome sequencing, and no off-target effects were detected at the whole-genome level. In addition, effective editing at the relaxed TTV PAM site was successfully detected. Finally, a multiplex genome editing system was constructed by using CRISPR/Mb2Cas12a system to simultaneously target seven sites in a single crRNA array with TTTV PAMs.

本研究构建了一套由CRISPR/Mb2Cas12a介导的棉花高效基因组编辑系统。首先，我们设计了三款搭载不同表达盒的载体，以验证其在棉花中的适用性。我们已分别成功获得无棉酚植株与叶片褪绿植株，证实三款载体均可在棉花中实现高效编辑。此外，通过全基因组测序分析，我们证实Mb2Cas12a具备极高的编辑精准度，全基因组水平未检测到脱靶效应。除此以外，我们成功在宽松型TTV原型间隔序列毗邻基元（Protospacer Adjacent Motif，简称PAM）位点上检测到有效编辑。最后，我们利用CRISPR/Mb2Cas12a系统构建了多重基因组编辑体系，可通过单条携带TTTV PAM的CRISPR RNA（crRNA）阵列同时靶向七个靶位点。