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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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Barbosa,Maria C.; Tovar,Francisco J.; Bressan-Nascimento,Suzete. |
Four species of Tephritoidea, three from genus Anastrepha: A. obliqua (Macquart), A. sororcula Zucchi and A. serpentina (Wiedemann), and one from genus Ceratitis, C. capitata (Wiedemann) were compared based on puparium morphology and application of the Heteroduplex Mobility Assay (HMA). Puparia were characterized for the first time using the spiracular posterior plate morphology. Application of HMA allowed the detection of variability in the D2 domain from 28S rRNA gene in all four species (confirmed by sequencing). This is a fast, simple, sensitive and inexpensive assay that was used for the first time in the analysis of two Tephritoidea genera. The detected variability suggests that the tecnique has great potential for rapid determination of infestations... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Fruit fly; Tephritoidea; Puparium; HMA. |
Ano: 2005 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1519-566X2005000600007 |
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