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

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 h...

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Published inActa tropica Vol. 178; pp. 190 - 195
Main Authors Romagnoli, Bruno A.A., Picchi, Gisele F.A., Hiraiwa, Priscila M., Borges, Beatriz S., Alves, Lysangela R., Goldenberg, Samuel
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.02.2018
Subjects
CRISPR-Cas Systems
CRISPR/Cas9
Essential genes
Genetic Engineering - methods
Improved technology
Knockout
Trypanosoma cruzi
Trypanosoma cruzi - genetics
Essential genes
Improved technology
CRISPR/Cas9
Knockout
Trypanosoma cruzi
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Summary: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.
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ISSN:0001-706X
1873-6254
DOI:10.1016/j.actatropica.2017.11.013