Use of the heteroduplex mobility assay and cell sorting to select genome sequences of the CCR5 gene in HEK 293T cells edited by transcription activator-like effector nucleases

Nerys-Junior, Arildo, Costa, Lendel C., Braga-Dias, Luciene P., Oliveira, Márcia, Rossi, Átila D., Cunha, Rodrigo Delvecchio da, Gonçalves, Gabriel S., Tanuri, Amilcar

Use of the heteroduplex mobility assay and cell sorting to select genome sequences of the CCR5 gene in HEK 293T cells edited by transcription activator-like effector nucleases

AUTOR(ES)

Nerys-Junior, Arildo, Costa, Lendel C., Braga-Dias, Luciene P., Oliveira, Márcia, Rossi, Átila D., Cunha, Rodrigo Delvecchio da, Gonçalves, Gabriel S., Tanuri, Amilcar

FONTE

Genet. Mol. Biol.

DATA DE PUBLICAÇÃO

2014

RESUMO

Engineered nucleases such as zinc finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN) are one of the most promising tools for modifying genomes. These site-specific enzymes cause double- strand breaks that allow gene disruption or gene insertion, thereby facilitating genetic manipulation. The major problem associated with this approach is the labor-intensive procedures required to screen and confirm the cellular modification by nucleases. In this work, we produced a TALEN that targets the human CCR5 gene and developed a heteroduplex mobility assay for HEK 293T cells to select positive colonies for sequencing. This approach provides a useful tool for the quick detection and easy assessment of nuclease activity.