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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 Genet. Mol. Biol.
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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Genetic improvement of horticultural crops mediated by CRISPR/Cas: a new horizon of possibilities Horticultura Brasileira
Koltun,Alessandra; Erpen-Dalla Corte,Lígia; Mertz-Henning,Liliane Marcia; Gonçalves,Leandro SA.
ABSTRACT The burden of the current global challenge involving food security lies in the need to improve crop production. In this regard, biotechnology stands out as an essential tool to generate plants able to cope with pests, diseases, and harsh climatic conditions, and more efficient in the use of natural resources. An advanced approach to create genetic variability in a precise and targeted way, the genome-editing technique CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR associated proteins), has drawn the attention of breeders. The genome editing CRISPR/Cas system relies on a guiding RNA that directs a nuclease to generate a double-strand break (DSB) at a target DNA, activating the cell repair systems and eventually leading...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Genome editing; Plant breeding; Biotechnology; Induced mutations; Non-transgenic.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-05362018000300290
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Future directions in breeding for disease resistance in aquaculture species R. Bras. Zootec.
Houston,Ross D..
ABSTRACT Infectious disease is a major constraint for all species produced via aquaculture. The majority of farmed fish and shellfish production is based on stocks with limited or no selective breeding. Since disease resistance is almost universally heritable, there is huge potential to select for improved resistance to key diseases. This short review discusses the current methods of breeding more resistant aquaculture stocks, with success stories and current bottlenecks highlighted. The current implementation of genomic selection in breeding for disease resistance and routes to wider-scale implementation and improvement in aquaculture are discussed. Future directions are highlighted, including the potential of genome editing tools for mapping causative...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Genome editing; Genomic selection; Selective breeding.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982017000600545
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