A scalable pipeline for highly effective genetic modification of a malaria parasite
Otto, Thomas D.
Quail, Michael A.
Rayner, Julian C.
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Computational Bioscience Research Center (CBRC)
Pathogen Genomics Laboratory
Online Publication Date2011-10-23
Print Publication Date2011-12
Permanent link to this recordhttp://hdl.handle.net/10754/325358
MetadataShow full item record
AbstractIn 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.
CitationPfander 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.
PubMed Central IDPMC3431185
- Recombination-mediated genetic engineering of Plasmodium berghei DNA.
- Authors: Pfander C, Anar B, Brochet M, Rayner JC, Billker O
- Issue date: 2013
- Expansion of experimental genetics approaches for Plasmodium berghei with versatile transfection vectors.
- Authors: Kooij TW, Rauch MM, Matuschewski K
- Issue date: 2012 Sep
- PlasmoGEM, a database supporting a community resource for large-scale experimental genetics in malaria parasites.
- Authors: Schwach F, Bushell E, Gomes AR, Anar B, Girling G, Herd C, Rayner JC, Billker O
- Issue date: 2015 Jan
- Multifunctional Involvement of a C2H2 Zinc Finger Protein (PbZfp) in Malaria Transmission, Histone Modification, and Susceptibility to DNA Damage Response.
- Authors: Gopalakrishnan AM, Aly ASI, Aravind L, Kumar N
- Issue date: 2017 Aug 29
- A genome-scale vector resource enables high-throughput reverse genetic screening in a malaria parasite.
- Authors: Gomes AR, Bushell E, Schwach F, Girling G, Anar B, Quail MA, Herd C, Pfander C, Modrzynska K, Rayner JC, Billker O
- Issue date: 2015 Mar 11