A scalable pipeline for highly effective genetic modification of a malaria parasite

Claudia Pfander, Burcu Anar, Frank Schwach, Thomas D. Otto, Mathieu Brochet, Katrin Volkmann, Michael A. Quail, Arnab Pain, Barry Rosen, William Skarnes, Julian C. Rayner, Oliver Billker*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

60 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1078-1084
Number of pages7
JournalNature Methods
Volume8
Issue number12
DOIs
Publication statusPublished - Dec 2011
Externally publishedYes

Fingerprint

Dive into the research topics of 'A scalable pipeline for highly effective genetic modification of a malaria parasite'. Together they form a unique fingerprint.

Cite this