RNA interference protocol to silence oncogenic drivers in leukemia cell lines

Mandy Beyer; Oliver H Krämer

doi:10.1016/j.xpro.2022.101512

RNA interference protocol to silence oncogenic drivers in leukemia cell lines

STAR Protoc. 2022 Sep 16;3(3):101512. doi: 10.1016/j.xpro.2022.101512. Epub 2022 Jul 1.

Authors

Mandy Beyer¹, Oliver H Krämer²

Affiliations

¹ Institute of Toxicology, Johannes-Gutenberg University of Mainz, Obere Zahlbacher St. 67, 55131 Mainz, Germany. Electronic address: manbeyer@uni-mainz.de.
² Institute of Toxicology, Johannes-Gutenberg University of Mainz, Obere Zahlbacher St. 67, 55131 Mainz, Germany. Electronic address: okraemer@uni-mainz.de.

Abstract

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).

Keywords: Cancer; Cell Biology; Cell culture; Cell-based Assays; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cats
Cell Line
Leukemia* / genetics
Protein-Tyrosine Kinases / genetics
RNA Interference
RNA, Small Interfering / genetics

Substances

RNA, Small Interfering
Protein-Tyrosine Kinases