| Autor: |
Dimitrios Papadopoulos, Carsten Patrick Ade, Martin Eilers |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
STAR Protocols, Vol 3, Iss 1, Pp 101183- (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2666-1667 |
| DOI: |
10.1016/j.xpro.2022.101183 |
| Popis: |
Summary: Here, we detail a protocol for the generation of pooled short hairpin RNA (shRNA) libraries. We cover the design of optimized miR-E backbone shRNAs, cloning into a Tet-on vector system, and transformation of competent bacteria. We also describe library quality check by next-generation sequencing, and finally the production of lentiviruses. This protocol will generate high-quality inducible libraries suitable for both genome-wide and targeted functional genomics screens, allowing the high-throughput interrogation of protein depletion effects in the cell system of choice.For complete details on the use and execution of this protocol, please refer to Papadopoulos et al. (2022). |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
|
