| Autor: |
Elisabeth, Pfeiffenberger, Reinhard, Sigl, Stephan, Geley |
| Rok vydání: |
2016 |
| Předmět: |
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| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 1448 |
| ISSN: |
1940-6029 |
| Popis: |
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). |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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