Conditional RNAi Using the Lentiviral GLTR System.
Methods Mol Biol
; 1448: 121-38, 2016.
Article
em En
| MEDLINE
| ID: mdl-27317178
ABSTRACT
RNA interference (RNAi) has become an essential technology for functional gene analysis. Its success depends on the effective expression of target gene-specific RNAi-inducing small double-stranded interfering RNA molecules (siRNAs). Here, were describe the use of a recently developed lentiviral RNAi system that allows the rapid generation of stable cell lines with inducible RNAi based on conditional expression of double-stranded short hairpin RNA (shRNA). These lentiviral vectors can be generated rapidly using the GATEWAY recombination cloning technology. Conditional cell lines can be established by using either a two-vector system in which the regulator is encoded by a separate vector or by a one-vector system. The available different lentiviral vectors for conditional shRNA expression cassette delivery co-express additional genes that allow (1) the use of fluorescent proteins for color-coded combinatorial RNAi or monitoring RNAi induction (pGLTR-FP), (2) selection of transduced cells (pGLTR-S), and (3) the generation of conditional cell lines using a one-vector system (pGLTR-X).
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Base de dados:
MEDLINE
Assunto principal:
Técnicas de Transferência de Genes
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Lentivirus
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Interferência de RNA
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Vetores Genéticos
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article