期刊
MOLECULAR AND BIOCHEMICAL PARASITOLOGY
卷 203, 期 1-2, 页码 25-33出版社
ELSEVIER
DOI: 10.1016/j.molbiopara.2015.10.004
关键词
Trypanosoma cruzi; Genome editing; Molecular parasitology
资金
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [590109/2011-1, 445456/2014-0, 305314/2013-0, 401355/2014-4, 309207/2014-2, 552636/2011-8]
- Fundacao de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG)
- Institute Nacional de Ciencia e Tecnologia de Vacinas (INCTV)
- CNPq-Brazil
- Coordenacao de Aperfeicoamento de Pessoal de Ensino Superior (CAPES)
Trypanosoma cruzi is a protozoan parasite that causes Chagas disease, an illness that affects 6-7 million people and for which there is no effective drug therapy or vaccine. The publication of its complete genome sequence allowed a rapid advance in molecular studies including in silico screening of genes involved with pathogenicity as well as molecular targets for the development of new diagnostic methods, drug therapies and prophylactic vaccines. Alongside with in silico genomic analyses, methods to study gene function in this parasite such as gene deletion, overexpression, mutant complementation and reporter gene expression have been largely explored. More recently, the use of genome-wide strategies is producing a shift towards a global perspective on gene function studies, with the examination of the expression and biological roles of gene networks in different stages of the parasite life cycle and under different contexts of host parasite interactions. Here we describe the molecular tools and protocols currently available to perform genetic manipulation of the T. cruzi genome, with emphasis on recently described strategies of gene editing that will facilitate large-scale functional genomic analyses. These new methodologies are long overdue, since more efficient protocols for genetic manipulation in T. cruzi are urgently needed for a better understanding of the biology of this parasite and molecular processes involved with the complex and often harmful, interaction with its human host. (C) 2015 Elsevier B.V. All rights reserved.
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