Journal
PARASITOLOGY
Volume 141, Issue 11, Pages 1390-1398Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S0031182014000559
Keywords
apicomplexan parasites; gold-standard genetic tools; conditional mutants; tetracycline inducible system; protein destabilization; DiCre
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Funding
- Wellcome Trust Senior Fellowship [087582/Z/08/Z]
- ERC Starting Grant [ERC-2012-StG 309255]
- Wellcome Trust [085349]
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The dissection of apicomplexan biology has been highly influenced by the genetic tools available for manipulation of parasite DNA. Here, we describe different techniques available for the generation of conditional mutants. Comparison of the advantages and disadvantages of the three most commonly used regulation systems: the tetracycline inducible system, the regulation of protein stability and site-specific recombination are discussed. Using some previously described examples we explore some of the pitfalls involved in gene-function analysis using these systems that can lead to wrong or over-interpretation of phenotypes. We will also mention different options to standardize the application of these techniques for the characterization of gene function in high-throughput.
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