A CRISPR/LbCas12a-based method for highly efficient multiplex gene editing in Physcomitrella patens
Pu, Xiaojun1,2; Liu, Lina1,2,3; Li, Ping1,2,3; Huo, Heqiang4; Dong, Xiumei1,2; Xie, Kabin5; Yang, Hong1,2,3; Liu, Li1,2
通讯作者Liu, Li(liulia@mail.kib.ac.cn)
2019-09-06
发表期刊PLANT JOURNAL
ISSN0960-7412
页码10
摘要Due to their high efficiency, specificity, and flexibility, programmable nucleases, such as those of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a (Cpf1) system, have greatly expanded the applicability of editing the genomes of various organisms. Genes from different gene families or genes with redundant functions in the same gene family can be examined by assembling multiple CRISPR RNAs (crRNAs) in a single vector. However, the activity and efficiency of CRISPR/Cas12a in the non-vascular plant Physcomitrella patens are largely unknown. Here, we demonstrate that LbCas12a together with its mature crRNA can target multiple loci simultaneously in P. patens with high efficiency via co-delivery of LbCas12a and a crRNA expression cassette in vivo. The mutation frequencies induced by CRISPR/LbCas12a at a single locus ranged from 26.5 to 100%, with diverse deletions being the most common type of mutation. Our method expands the repertoire of genome editing tools available for P. patens and facilitates the creation of loss-of-function mutants of multiple genes from different gene families.
关键词CRISPR Cas12a CRISPR Cas9 genome editing Physcomitrella patens technical advance
DOI10.1111/tpj.14478
收录类别SCI
语种英语
WOS记录号WOS:000485845800001
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文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/68705
专题资源植物与生物技术所级重点实验室
通讯作者Liu, Li
作者单位1.Chinese Acad Sci, Kunming Inst Bot, Key Lab Econ Plants & Biotechnol, Kunming 650201, Yunnan, Peoples R China
2.Yunnan Key Lab Wild Plant Resources, Kunming 650201, Yunnan, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Univ Florida, Dept Environm Hort, Mid Florida Res & Educ Ctr, Gainesville, FL 32703 USA
5.Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Wuhan 430070, Hubei, Peoples R China
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Pu, Xiaojun,Liu, Lina,Li, Ping,et al. A CRISPR/LbCas12a-based method for highly efficient multiplex gene editing in Physcomitrella patens[J]. PLANT JOURNAL,2019:10.
APA Pu, Xiaojun.,Liu, Lina.,Li, Ping.,Huo, Heqiang.,Dong, Xiumei.,...&Liu, Li.(2019).A CRISPR/LbCas12a-based method for highly efficient multiplex gene editing in Physcomitrella patens.PLANT JOURNAL,10.
MLA Pu, Xiaojun,et al."A CRISPR/LbCas12a-based method for highly efficient multiplex gene editing in Physcomitrella patens".PLANT JOURNAL (2019):10.
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