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. 2018 Feb:178:190-195.
doi: 10.1016/j.actatropica.2017.11.013. Epub 2017 Nov 22.

Improvements in the CRISPR/Cas9 system for high efficiency gene disruption in Trypanosoma cruzi

Bruno A A Romagnoli  1 , Gisele F A Picchi  2 , Priscila M Hiraiwa  3 , Beatriz S Borges  4 , Lysangela R Alves  5 , Samuel Goldenberg  6
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Free article

Improvements in the CRISPR/Cas9 system for high efficiency gene disruption in Trypanosoma cruzi

Bruno A A Romagnoli et al. Acta Trop. 2018 Feb.
Free article

Abstract

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, affects millions of individuals around the world. Although it has been known for more than a century, the study of T. cruzi has been a challenge, particularly due to the scarcity of tools for genome inquiries. Recently, strategies have been described allowing gene disruption in T. cruzi by the CRISPR/Cas9 nuclease system. Although these strategies demonstrated success in deleting some genes, several aspects could be improved to increase the efficiency of the CRISPR/Cas9 system in T. cruzi. Here, we report a strategy, based on adaptations and improvements of the two previously described systems, that results in efficient gene disruption that can be applied to any target, including the study of essential genes.

Keywords: CRISPR/Cas9; Essential genes; Improved technology; Knockout; Trypanosoma cruzi.

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