A scalable pipeline for highly effective genetic modification of a malaria parasite
Type
ArticleAuthors
Pfander, ClaudiaAnar, Burcu
Schwach, Frank
Otto, Thomas D.
Brochet, Mathieu
Volkmann, Katrin
Quail, Michael A.
Pain, Arnab

Rosen, Barry
Skarnes, William
Rayner, Julian C.
Billker, Oliver
KAUST Department
Biological and Environmental Sciences and Engineering (BESE) DivisionBioscience Program
Computational Bioscience Research Center (CBRC)
Pathogen Genomics Laboratory
Date
2011-10-23Online Publication Date
2011-10-23Print Publication Date
2011-12Permanent link to this record
http://hdl.handle.net/10754/325358
Metadata
Show full item recordAbstract
In malaria parasites, the systematic experimental validation of drug and vaccine targets by reverse genetics is constrained by the inefficiency of homologous recombination and by the difficulty of manipulating adenine and thymine (A+T)-rich DNA of most Plasmodium species in Escherichia coli. We overcame these roadblocks by creating a high-integrity library of Plasmodium berghei genomic DNA (>77% A+T content) in a bacteriophage N15-based vector that can be modified efficiently using the lambda Red method of recombineering. We built a pipeline for generating P. berghei genetic modification vectors at genome scale in serial liquid cultures on 96-well plates. Vectors have long homology arms, which increase recombination frequency up to tenfold over conventional designs. The feasibility of efficient genetic modification at scale will stimulate collaborative, genome-wide knockout and tagging programs for P. berghei. © 2011 Nature America, Inc. All rights reserved.Citation
Pfander C, Anar B, Schwach F, Otto TD, Brochet M, et al. (2011) A scalable pipeline for highly effective genetic modification of a malaria parasite. Nature Methods 8: 1078-1082. doi:10.1038/nmeth.1742.Publisher
Springer NatureJournal
Nature MethodsPubMed ID
22020067PubMed Central ID
PMC3431185ae974a485f413a2113503eed53cd6c53
10.1038/nmeth.1742
Scopus Count
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