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RNA interference protocol to silence oncogenic drivers in leukemia cell lines

Genetic silencing of leukemia-associated proteins with small interfering RNAs (siRNAs) is a straightforward way to delineate their functions. It can be very challenging to deliver siRNAs to leukemia-derived cells with high transfection efficiency and without compromising their viability. This protoc...

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Detalles Bibliográficos
Autores principales: Beyer, Mandy, Krämer, Oliver H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9254495/
https://www.ncbi.nlm.nih.gov/pubmed/35779262
http://dx.doi.org/10.1016/j.xpro.2022.101512
Descripción
Sumario:Genetic silencing of leukemia-associated proteins with small interfering RNAs (siRNAs) is a straightforward way to delineate their functions. It can be very challenging to deliver siRNAs to leukemia-derived cells with high transfection efficiency and without compromising their viability. This protocol describes an efficient approach to silence oncogenic feline McDonough sarcoma (FMS)-like tyrosine kinase-3 in leukemia cells using siRNAs that are delivered by electroporation. The protocol maintains high cell viability and is generally useful to decrease RNAs encoding proteins of interest. For complete details on the use and execution of this protocol, please refer to Beyer et al. (2022).