CRISPR/Cas9 Mutagenesis and Expression of Dominant Mutant Transgenes as Functional Genomic Approaches in Parasitic Nematodes.

Frontiers in Genetics
James B Lok

Abstract

DNA transformation of parasitic nematodes enables novel approaches to validating predictions from genomic and transcriptomic studies of these important pathogens. Notably, proof of principle for CRISPR/Cas9 mutagenesis has been achieved in Strongyloides spp., allowing identification of molecules essential to the functions of sensory neurons that mediate behaviors comprising host finding, invasion, and location of predilection sites by parasitic nematodes. Likewise, CRISPR/Cas9 knockout of the developmental regulatory transcription factor Ss-daf-16 has validated its function in regulating morphogenesis of infective third-stage larvae in Strongyloides stercoralis. While encouraging, these studies underscore challenges that remain in achieving straightforward validation of essential intervention targets in parasitic nematodes. Chief among these is the likelihood that knockout of multifunctional regulators like Ss-DAF-16 or its downstream mediator, the nuclear receptor Ss-DAF-12, will produce phenotypes so complex as to defy interpretation and will render affected worms incapable of infecting their hosts, thus preventing establishment of stable mutant lines. Approaches to overcoming these impediments could involve refinements to cu...Continue Reading

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Citations

Sep 1, 2020·PLoS Neglected Tropical Diseases·Canhui LiuThomas R Unnasch
Jan 14, 2020·Current Tropical Medicine Reports·Tegegn G Jaleta, James B Lok
May 13, 2020·Biology Methods and Protocols·Elizabeth CulpBhagwati P Gupta
Nov 30, 2020·International Journal for Parasitology. Drugs and Drug Resistance·Steffen R HahnelDaniel Kulke
Jun 8, 2021·Trends in Parasitology·Mostafa Zamanian, John D Chan

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Methods Mentioned

BETA
transgenic
transfection
transfecting
genotyping
PCR

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