Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment
| Autor: | Geethu E. Thomas, Ayesh K. Seneviratne, Rose Hurren, Neil MacLean, Aaron D. Schimmer, Rashim Pal Singh |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Genetic Vectors
Biology General Biochemistry Genetics and Molecular Biology Viral vector Small hairpin RNA 03 medical and health sciences Transduction (genetics) Mice 0302 clinical medicine In vivo Transduction Genetic Protocol Tumor Cells Cultured In vitro study Animals Humans lcsh:Science (General) Gene 030304 developmental biology Cancer 0303 health sciences General Immunology and Microbiology General Neuroscience Stem Cells Lentivirus Myeloid leukemia 3. Good health Leukemia Myeloid Acute Cancer research Stem cell 030217 neurology & neurosurgery Neoplasm Transplantation lcsh:Q1-390 |
| Zdroj: | STAR Protocols STAR Protocols, Vol 1, Iss 3, Pp 100163-(2020) |
| ISSN: | 2666-1667 |
| Popis: | Summary We describe a method to silence genes in primary acute myeloid leukemia cells by transducing them with shRNA in lentiviral vectors. The transduction of primary non-adherent cells is particularly challenging. The protocol will aid in performing such experiments and is particularly helpful to prepare cells for in vivo engraftment studies. Use of a special medium supplemented with cytokines preserves the viability of the leukemic stem cells and their ability to engraft the marrow of immune-deficient mice. For complete details on the use and execution of this protocol, please refer to Singh et al. (2020). Graphical Abstract Highlights • An optimized protocol to silence genes in primary AML cells • Transduces primary AML cells with shRNA in lentiviral vectors • Preserves viability of the primary cells • Suitable for downstream assays including cell viability and in vivo engraftment We describe a method to silence genes in primary acute myeloid leukemia cells by transducing them with shRNA in lentiviral vectors. The transduction of primary non-adherent cells is particularly challenging. The protocol will aid in performing such experiments and is particularly helpful to prepare cells for in vivo engraftment studies. Use of a special medium supplemented with cytokines preserves the viability of the leukemic stem cells and their ability to engraft the marrow of immune-deficient mice. |
| Databáze: | OpenAIRE |
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