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A Cas9 transgenic Plasmodium yoelii parasite for efficient gene editing.

Authors :
Qian, Pengge
Wang, Xu
Yang, Zhenke
Li, Zhenkui
Gao, Han
Su, Xin-zhuan
Cui, Huiting
Yuan, Jing
Source :
Molecular & Biochemical Parasitology. Jun2018, Vol. 222, p21-28. 8p.
Publication Year :
2018

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 the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for genome editing in the parasites only introduced with cas9 plasmid. To establish the endogenous and constitutive expression of Cas9 protein in the rodent malaria parasite P. yoelii , we replaced the coding region of an endogenous gene sera1 with the intact SpCas9 coding sequence using the CRISPR/Cas9-mediated genome editing method, generating the cas9-knockin parasite ( PyCas9ki ) of the rodent malaria parasite P. yoelii . The resulted PyCas9ki parasite displays normal progression during the whole life cycle and possesses the Cas9 protein expression in asexual blood stage. By introducing the plasmid (pYCs) containing only sgRNA and homologous template elements, we successfully achieved both deletion and tagging modifications for different endogenous genes in the genome of PyCas9ki parasite. This cas9-knockin PyCas9ki parasite provides a new platform facilitating gene functions study in the rodent malaria parasite P. yoelii . [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
01666851
Volume :
222
Database :
Academic Search Index
Journal :
Molecular & Biochemical Parasitology
Publication Type :
Academic Journal
Accession number :
129946973
Full Text :
https://doi.org/10.1016/j.molbiopara.2018.04.003