Chapter title |
Conditional RNAi Using the Lentiviral GLTR System.
|
---|---|
Chapter number | 10 |
Book title |
Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3753-0_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3751-6, 978-1-4939-3753-0
|
Authors |
Elisabeth Pfeiffenberger, Reinhard Sigl, Stephan Geley |
Editors |
Maurizio Federico |
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). |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 2 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 2 | 100% |
Readers by discipline | Count | As % |
---|---|---|
Neuroscience | 2 | 100% |