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Nat Methods. 2014 Sep;11(9):915-8. doi: 10.1038/nmeth.3063. Epub 2014 Aug 10.

Efficient CRISPR-Cas9-mediated genome editing in Plasmodium falciparum.

Author information

1
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
1] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
3
1] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. [3] McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [4] Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

Abstract

Malaria is a major cause of global morbidity and mortality, and new strategies for treating and preventing this disease are needed. Here we show that the Streptococcus pyogenes Cas9 DNA endonuclease and single guide RNAs (sgRNAs) produced using T7 RNA polymerase (T7 RNAP) efficiently edit the Plasmodium falciparum genome. Targeting the genes encoding native knob-associated histidine-rich protein (kahrp) and erythrocyte binding antigen 175 (eba-175), we achieved high (≥ 50-100%) gene disruption frequencies within the usual time frame for generating transgenic parasites.

PMID:
25108687
PMCID:
PMC4199390
DOI:
10.1038/nmeth.3063
[Indexed for MEDLINE]
Free PMC Article

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