A Cas9 transgenic Plasmodium yoelii parasite for efficient gene editing.
作者： Pengge Qian , Xu Wang , Zhenke Yang , Zhenkui Li , Han Gao
DOI: 10.1016/J.MOLBIOPARA.2018.04.003
关键词: Cas9 、 Parasite hosting 、 Coding region 、 Gene 、 Biology 、 Genome 、 Genetics 、 Plasmodium yoelii 、 CRISPR 、 Genome editing
摘要: Abstract The RNA-guided endonuclease Cas9 has applied as an efficient gene-editing method in malaria parasite Plasmodium . However, the size (4.2 kb) of commonly used from Streptococcus pyogenes (SpCas9) limits its utility for genome editing parasites only introduced with cas9 plasmid. To establish endogenous and constitutive expression protein rodent P. yoelii , we replaced coding region gene sera1 intact SpCas9 sequence using CRISPR/Cas9-mediated method, generating cas9-knockin ( PyCas9ki ) resulted displays normal progression during whole life cycle possesses asexual blood stage. By introducing plasmid (pYCs) containing sgRNA homologous template elements, successfully achieved both deletion tagging modifications different genes parasite. This provides a new platform facilitating functions study
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