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| ROCHA-MARTINS,MAURÍCIO; CAVALHEIRO,GABRIEL R.; MATOS-RODRIGUES,GABRIEL E.; MARTINS,RODRIGO A.P.. |
| ABSTRACTGenome modification technologies are powerful tools for molecular biology and related areas. Advances in animal transgenesis and genome editing technologies during the past three decades allowed systematic interrogation of gene function that can help model how the genome influences cellular physiology. Genetic engineering via homologous recombination (HR) has been the standard method to modify genomic sequences. Nevertheless, nuclease-guided genome editing methods that were developed recently, such as ZFN, TALEN and CRISPR/Cas, opened new perspectives for biomedical research. Here, we present a brief historical perspective of genome modification methods, focusing on transgenic mice models. Moreover, we describe how new techniques were discovered... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Homologous recombination; DNA repair; Cre-LoxP; ZFN; TALEN; CRISPR/Cas9.. |
| Ano: 2015 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000301323 |
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| Nerys-Junior,Arildo; Costa,Lendel C.; Braga-Dias,Luciene P.; Oliveira,Márcia; Rossi,Átila D.; Cunha,Rodrigo Delvecchio da; Gonçalves,Gabriel S.; Tanuri,Amilcar. |
| 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. |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: CCR5; Genome editing; Gene knockout; TALEN; HMA. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572014000100018 |
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