Forward RNAi screens in human hematopoietic stem cells
(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.
(Less)
- author
- Galeev, Roman LU ; Karlsson, Christine LU ; Baudet, Aurélie LU and Larsson, Jonas LU
- organization
- publishing date
- 2017
- 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
- host publication
- Methods in Molecular Biology
- series title
- Methods in Molecular Biology
- volume
- 1622
- pages
- 22 pages
- publisher
- Humana Press
- external identifiers
-
- scopus:85021743188
- pmid:28674799
- 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
- 2025-01-07 17:26:09
@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}},
booktitle = {{Methods in Molecular Biology}},
issn = {{10643745}},
keywords = {{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}},
doi = {{10.1007/978-1-4939-7108-4_3}},
volume = {{1622}},
year = {{2017}},
}