RNAi screening of leukemia cells using electroporation

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

RNAi-mediated screening has been an integral tool for biological discovery for the past 15 years. A variety of approaches have been employed for implementation of this technique, including pooled, depletion/ enrichment screening with lentiviral shRNAs, and segregated screening of panels of individual siRNAs. The latter approach of siRNA panel screening requires efficient methods for transfection of siRNAs into the target cells. In the case of suspension leukemia cell lines and primary cells, many of the conventional transfection techniques using liposomal or calcium phosphate-mediated transfection provide very low efficiency. In this case, electroporation is the only transfection technique offering high efficiency transfection of siRNAs into the target leukemia cells. Here, we describe methods for optimization and implementation of siRNA electroporation into leukemia cell lines and primary patient specimens, and we further offer suggested electroporation settings for some commonly used leukemia cell lines.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages85-94
Number of pages10
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1470
ISSN (Print)1064-3745

Keywords

  • Electroporation
  • Leukemia
  • siRNA screening

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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