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| Pessoa,Luciana Santos; Vidal,Luãnna Liebscher; Costa,Emmerson C.B. da; Abreu,Celina Monteiro; Cunha,Rodrigo Delvecchio da; Valadão,Ana Luiza Chaves; Santos,André Felipe dos; Tanuri,Amilcar. |
| Abstract Approximately 185 million people worldwide are chronically infected with hepatitis C virus (HCV). The first-wave of approved NS3 protease inhibitors (PIs) were Telaprevir and Boceprevir, which are currently discontinued. Simeprevir is a second-wave PI incorporated into the Brazilian hepatitis C treatment protocol. Drug resistance plays a key role in patients' treatment regimen. Here, we developed a simple phenotypic assay to evaluate the impact of resistance mutations in HCV NS3 protease to PIs, using a protein expression vector containing wild type NS3 protease domain and NS4A co-factor. We analyzed the impact of five resistance mutations (T54A, V36M, V158I, V170I and T54S+V170I) against Telaprevir, Boceprevir and Simeprevir. Protein... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Protease inhibitor; Phenotypic test; Hepatitis C virus; HCV NS3 protease; Resistance mutation. |
| Ano: 2016 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000300358 |
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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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