title: Editing the Plasmodium vivax genome, using zinc-finger nucleases.
abstract: Plasmodium vivax is a major cause of malaria morbidity worldwide yet has remained genetically intractable. To stably modify this organism, we used zinc-finger nucleases (ZFNs), which take advantage of homology-directed DNA repair mechanisms at the site of nuclease action. Using ZFNs specific to the gene encoding P. vivax dihydrofolate reductase (pvdhfr), we transfected blood specimens from Saimiri boliviensis monkeys infected with the pyrimethamine (Pyr)-susceptible Chesson strain with a ZFN plasmid carrying a Pyr-resistant mutant pvdhfr sequence. We obtained Pyr-resistant parasites in vivo that carried mutant pvdhfr and additional silent mutations designed to confirm editing. These results herald the era of stable P. vivax genetic modifications.
authors.affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Roberto R
authors.initials: RR
authors.lastname: Moraes Barros
authors.affiliation: Department of Microbiology and Immunology.
authors.forename: Judith
authors.initials: J
authors.lastname: Straimer
authors.affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Juliana M
authors.initials: JM
authors.lastname: Sa
authors.affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Salzman
authors.affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Viviana A
authors.initials: VA
authors.lastname: Melendez-Muniz
authors.affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Jianbing
authors.initials: J
authors.lastname: Mu
authors.affiliation: Department of Microbiology and Immunology Division of Infectious Diseases, Department of Medicine, Columbia University College of Physicians and Surgeons, New York, New York.
authors.forename: David A
authors.initials: DA
authors.lastname: Fidock
authors.affiliation: Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Wellems
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology; D003851:Deoxyribonucleases / Q000235:genetics*; D004351:Drug Resistance; D017125:Genes, Protozoan / Q000235:genetics; D018503:Genome, Protozoan; D016780:Malaria, Vivax / Q000188:drug therapy / Q000469:parasitology; D010966:Plasmodium vivax / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics; D011739:Pyrimethamine / Q000494:pharmacology; D012453:Saimiri / Q000469:parasitology; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D016335:Zinc Fingers
chemical_list: D000962:Antimalarials; D015800:Protozoan Proteins; D013762:Tetrahydrofolate Dehydrogenase; D003851:Deoxyribonucleases; D011739:Pyrimethamine
keywords: Saimiri boliviensis; allelic modification; dihydrofolate reductase-thymidylate synthase; malaria; pyrimethamine; transfection
references.citation: Trends Parasitol. 2011 Jan;27(1):31-9
references.pmid: 20663715
references.citation: Mol Biochem Parasitol. 2006 Sep;149(1):99-101
references.pmid: 16716418
references.citation: J Med Chem. 2013 Oct 24;56(20):7761-71
references.pmid: 23927658
references.citation: PLoS One. 2012;7(7):e40416
references.pmid: 22792308
references.citation: Antimicrob Agents Chemother. 2002 Dec;46(12):3947-53
references.pmid: 12435700
references.citation: Nat Med. 2013 Feb;19(2):156-67
references.pmid: 23389616
references.citation: Nature. 2013 Dec 12;504(7479):248-253
references.pmid: 24284631
references.citation: Nat Rev Microbiol. 2013 Oct;11(10):701-12
references.pmid: 24037451
references.citation: Nat Methods. 2012 Oct;9(10):993-8
references.pmid: 22922501
references.citation: Mol Microbiol. 2008 Jun;68(6):1535-46
references.pmid: 18410493
references.citation: Antimicrob Agents Chemother. 2005 Dec;49(12):4928-33
references.pmid: 16304154
references.citation: Trends Biotechnol. 2013 Jul;31(7):397-405
references.pmid: 23664777
references.citation: Nat Methods. 2011 Jan;8(1):74-9
references.pmid: 21131970
references.citation: Cell Microbiol. 2014 May;16(5):612-20
references.pmid: 24506567
references.citation: Am J Trop Med Hyg. 1978 Jul;27(4):703-17
references.pmid: 99056