Taming Parasites by Tailoring Them

Abstract

The next-generation gene editing based on CRISPR (clustered regularly interspaced short palindromic repeats) has been successfully implemented in a wide range of organisms including some protozoan parasites. However, application of such a versatile game-changing technology in molecular parasitology remains fairly underexplored. Here, we briefly introduce state-of-the-art in human and mouse research and usher new directions to drive the parasitology research in the years to come. In precise, we outline contemporary ways to embolden existing apicomplexan and kinetoplastid parasite models by commissioning front-line gene-tailoring methods, and illustrate how we can break the enduring gridlock of gene manipulation in non-model parasitic protists to tackle intriguing questions that remain long unresolved otherwise. We show how a judicious solicitation of the CRISPR technology can eventually balance out the two facets of pathogen-host interplay.

Robust and stable transcriptional repression inGiardiausing CRISPRi

“…Molecular genetic tool development in Giardia has focused on transient translational repression by electroporation of morpholinos (Carpenter and Cande, 2009) or on the overexpression of long double-stranded RNAs or hammerhead ribozymes for transcriptional repression (Dan et al , 2000; Chen et al , 2007). While CRISPR/Cas9-mediated knockout strategies have recently been used for genome engineering in several parasitic protists (Ren and Gupta, 2017), this is the first demonstration of CRISPRi-mediated transcriptional repression in these organisms.…”

Protean permeases: Diverse roles for membrane transport proteins in kinetoplastid protozoa

“…Since a major function of transporters is to allow the influx of nutrients from host to parasite, I will discuss a few of the permeases that mediate uptake of metabolically important solutes. The advent of gene knockout technology, now facilitated by the recent introduction of CRISPR-Cas9 mediated gene editing [4, 5] and other sophisticated molecular genetic methodologies [6], provides a powerful strategy for addressing the functions of individual transporters and their variants. In addition, gene knockout and RNAi approaches allow investigators to determine whether specific permeases are essential for parasite viability, thus identifying those that could be targets for development of novel drugs.…”

CRISPR-based genomic tools for the manipulation of genetically intractable microorganisms

“…As with microbial fungi, clinically important eukaryotic parasites, including Trypanosoma cruzi , Toxoplasma gondii , Leishmania donovani , Cryptosporidium parvum, and the malarial parasite Plasmodium falciparum , have historically had limited genetic tools available for the facile generation of genetic mutants 25,26,41 ; yet new applications of CRISPR technology, are enabling efficient genetic disruptions in these parasites 24,[42][43][44][45][46] . An impressive use of CRISPR technology, was applied in the apicomplexan parasite T. gondii 42 -a ubiquitous parasite that can cause devastating congenital disease or fetal death if transmitted from mother to fetus.…”