Forward RNAi screens in human hematopoietic stem cells

Research output: Chapter in Book/Report/Conference proceedingBook chapter

Abstract

Identifying the genes and pathways that regulate self-renewal and differentiation in somatic stem cells is a central goal in stem cell and cancer biology. Here, we describe a method for RNA interference (RNAi)-based screens combined with next-generation sequencing (NGS) in primary human hematopoietic stem and progenitor cells (HSPCs). These cells are suitable targets for complex, selection-based screens using pooled lentiviral short hairpin RNA (shRNA) libraries. The screening approach presented in this chapter is a promising tool to dissect regulatory mechanisms in hematopoietic stem cells (HSCs) and somatic stem cells in general, and may be particularly useful to identify gene targets and modifiers that can be further exploited in strategies for ex vivo stem cell expansion.

Details

Authors
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell and Molecular Biology

Keywords

  • Forward genetic screens, Functional validation, Hematopoietic stem cells, Next-generation sequencing, Positive selection, RNA interference
Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages29-50
Number of pages22
Volume1622
Publication statusPublished - 2017
Publication categoryResearch
Peer-reviewedYes

Publication series

NameMethods in Molecular Biology
Volume1622
ISSN (Print)10643745