Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/28639
Title: Comparison of the editing patterns and editing efficiencies of TALEN and CRISPR-Cas9 when targeting the human CCR5 gene
Authors: Nerys Junior, Arildo
Dias, Luciene P. Braga
Pezzuto, Paula
Almeida, Vinícius Cotta de
Tanuri, Amilcar
Affilliation: Universidade Federal do Rio de Janeiro. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Ciência e Tecnologia em Biomodelos. Laboratório do Serviço de Biotecnologia e Desenvolvimento Animal. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Laboratório de Virologia Molecular. Rio de Janeiro, RJ, Brasil.
Abstract: The human C-C chemokine receptor type-5 (CCR5) is the major transmembrane co-receptor that mediates HIV-1 entry into target CD4+ cells. Gene therapy to knock-out the CCR5 gene has shown encouraging results in providing a functional cure for HIV-1 infection. In gene therapy strategies, the initial region of the CCR5 gene is a hotspot for producing functional gene knock-out. Such target gene editing can be done using programmable endonucleases such as transcription activator-like effector nucleases (TALEN) or clustered regularly interspaced short palindromic repeats (CRISPR-Cas9). These two gene editing approaches are the most modern and effective tools for precise gene modification. However, little is known of potential differences in the efficiencies of TALEN and CRISPR-Cas9 for editing the beginning of the CCR5 gene. To examine which of these two methods is best for gene therapy, we compared the patterns and amount of editing at the beginning of the CCR5 gene using TALEN and CRISPR-Cas9 followed by DNA sequencing. This comparison revealed that CRISPR-Cas9 mediated the sorting of cells that contained 4.8 times more gene editing than TALEN+ transfected cells.
Keywords: CCR5
CRISPR-Cas9
efficiency
gene editing
TALEN
DeCS: Nucleases dos Efetores Semelhantes a Ativadores de Transcrição
Sistemas CRISPR-Cas
Receptores CCR5
Eficiência
Issue Date: 2018
Publisher: Sociedade Brasileira de Genética
Citation: NERYS JUNIOR, Arildo; et al. Comparison of the editing patterns and editing efficiencies of TALEN and CRISPR-Cas9 when targeting the human CCR5 gene. Genetics and Molecular Biology, v.41, n.1, p.167-179, 2018.
DOI: 10.1590/1678-4685-GMB-2017-0065
ISSN: 1415-4757
Copyright: open access
Appears in Collections:IOC - Artigos de Periódicos

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