Chapter title |
Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAs
|
---|---|
Chapter number | 17 |
Book title |
RNAi and Small Regulatory RNAs in Stem Cells
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7108-4_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7106-0, 978-1-4939-7108-4
|
Authors |
Wang, Jianlong, Jianlong Wang |
Editors |
Baohong Zhang |
Abstract |
RNA interference (RNAi) is a powerful gene knockdown technology that has been applied for functional genetic loss-of-function studies in many model eukaryotic systems, including embryonic stem cells (ESCs). Application of RNAi in ESCs allows for dissection of mechanisms by which ESCs self-renew and maintain pluripotency and also for specifying particular cell types needed for cell replacement therapies. Potent RNAi response can be induced by expression of a microRNA-embedded short-hairpin RNA (shRNA(mir)) cassette that is integrated in the genome by virus infection or site-specific recombination at a defined locus. In this chapter, I will provide detailed protocols to perform shRNA(mir)-mediated RNAi studies in mouse ESCs using retrovirus infection and loxP site-directed recombination for efficient constitutive and inducible gene knockdown, respectively. |
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Student > Master | 2 | 29% |
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