STAR Protocols (Dec 2020)
Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment
Abstract
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).