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Forward RNAi screens in human hematopoietic stem cells

Galeev, Roman LU ; Karlsson, Christine LU ; Baudet, Aurélie LU and Larsson, Jonas LU (2017) In Methods in Molecular Biology 1622. p.29-50
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... (More)

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.

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Please use this url to cite or link to this publication:
author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Forward genetic screens, Functional validation, Hematopoietic stem cells, Next-generation sequencing, Positive selection, RNA interference
in
Methods in Molecular Biology
volume
1622
pages
22 pages
publisher
Humana Press
external identifiers
  • scopus:85021743188
ISSN
10643745
DOI
10.1007/978-1-4939-7108-4_3
language
English
LU publication?
yes
id
5938c7c0-1998-4cba-9255-d388fe336773
date added to LUP
2017-07-19 14:53:34
date last changed
2017-07-20 03:00:02
@inbook{5938c7c0-1998-4cba-9255-d388fe336773,
  abstract     = {<p>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.</p>},
  author       = {Galeev, Roman and Karlsson, Christine and Baudet, Aurélie and Larsson, Jonas},
  issn         = {10643745},
  keyword      = {Forward genetic screens,Functional validation,Hematopoietic stem cells,Next-generation sequencing,Positive selection,RNA interference},
  language     = {eng},
  pages        = {29--50},
  publisher    = {Humana Press},
  series       = {Methods in Molecular Biology},
  title        = {Forward RNAi screens in human hematopoietic stem cells},
  url          = {http://dx.doi.org/10.1007/978-1-4939-7108-4_3},
  volume       = {1622},
  year         = {2017},
}