Site-specific genome editing in Plasmodium falciparum using engineered zinc-finger nucleases

Judith Straimer, Marcus C.S. Lee, Andrew H. Lee, Bryan Zeitler, April E. Williams, Jocelynn R. Pearl, Lei Zhang, Edward J. Rebar, Philip D. Gregory, Manuel Llinás, Fyodor D. Urnov, David A. Fidock

Research output: Contribution to journalArticlepeer-review

115 Scopus citations

Abstract

Malaria afflicts over 200 million people worldwide, and its most lethal etiologic agent, Plasmodium falciparum, is evolving to resist even the latest-generation therapeutics. Efficient tools for genome-directed investigations of P. falciparum-induced pathogenesis, including drug-resistance mechanisms, are clearly required. Here we report rapid and targeted genetic engineering of this parasite using zinc-finger nucleases (ZFNs) that produce a double-strand break in a user-defined locus and trigger homology-directed repair. Targeting an integrated egfp locus, we obtained gene-deletion parasites with unprecedented speed (2 weeks), both with and without direct selection. ZFNs engineered against the parasite gene pfcrt, responsible for escape under chloroquine treatment, rapidly produced parasites that carried either an allelic replacement or a panel of specified point mutations. This method will enable a diverse array of genome-editing approaches to interrogate this human pathogen.

Original languageEnglish (US)
Pages (from-to)993-998
Number of pages6
JournalNature methods
Volume9
Issue number10
DOIs
StatePublished - Oct 2012

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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