Arrayed molecular barcoding of leukemic stem cells
(2021) In Methods in Molecular Biology 2185. p.345-359- Abstract
Functional screens on cancer cells using compound or protein libraries are usually performed in vitro. However, to assess the effects on leukemia stem cells (LSCs) in a screening setting, methodologies that allow for a high-throughput in vivo readout of leukemia-initiating activity are needed. One experimental approach to solve this issue is to genetically label, also referred to as “barcoding,” the leukemia cells in an arrayed format prior to exposing them to separate experimental conditions. The cells can then be pooled and injected into mice for competitive readout of leukemia-initiating activity. Here, we describe a procedure for combining lentiviral arrayed molecular barcoding of leukemia cells with next-generation sequencing, to... (More)
Functional screens on cancer cells using compound or protein libraries are usually performed in vitro. However, to assess the effects on leukemia stem cells (LSCs) in a screening setting, methodologies that allow for a high-throughput in vivo readout of leukemia-initiating activity are needed. One experimental approach to solve this issue is to genetically label, also referred to as “barcoding,” the leukemia cells in an arrayed format prior to exposing them to separate experimental conditions. The cells can then be pooled and injected into mice for competitive readout of leukemia-initiating activity. Here, we describe a procedure for combining lentiviral arrayed molecular barcoding of leukemia cells with next-generation sequencing, to enable screens on leukemia cells ex vivo followed by an in vivo competitive readout of LSC function. This methodology can also be applied to other model systems in which a competitive in vivo readout of cells is needed.
(Less)
- author
- Chapellier, Marion LU and Järås, Marcus LU
- organization
- publishing date
- 2021
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- keywords
- Arrayed screens, In vivo competition, Leukemic stem cells, Molecular barcoding, Next-generation sequencing
- host publication
- Leukemia Stem Cells
- series title
- Methods in Molecular Biology
- volume
- 2185
- pages
- 15 pages
- publisher
- Humana Press
- external identifiers
-
- pmid:33165859
- scopus:85095963680
- ISSN
- 1064-3745
- 1940-6029
- DOI
- 10.1007/978-1-0716-0810-4_21
- language
- English
- LU publication?
- yes
- id
- cf594e51-60ec-48bf-ae0a-d6c466bed354
- date added to LUP
- 2020-12-08 12:58:16
- date last changed
- 2025-04-04 15:05:58
@inbook{cf594e51-60ec-48bf-ae0a-d6c466bed354,
abstract = {{<p>Functional screens on cancer cells using compound or protein libraries are usually performed in vitro. However, to assess the effects on leukemia stem cells (LSCs) in a screening setting, methodologies that allow for a high-throughput in vivo readout of leukemia-initiating activity are needed. One experimental approach to solve this issue is to genetically label, also referred to as “barcoding,” the leukemia cells in an arrayed format prior to exposing them to separate experimental conditions. The cells can then be pooled and injected into mice for competitive readout of leukemia-initiating activity. Here, we describe a procedure for combining lentiviral arrayed molecular barcoding of leukemia cells with next-generation sequencing, to enable screens on leukemia cells ex vivo followed by an in vivo competitive readout of LSC function. This methodology can also be applied to other model systems in which a competitive in vivo readout of cells is needed.</p>}},
author = {{Chapellier, Marion and Järås, Marcus}},
booktitle = {{Leukemia Stem Cells}},
issn = {{1064-3745}},
keywords = {{Arrayed screens; In vivo competition; Leukemic stem cells; Molecular barcoding; Next-generation sequencing}},
language = {{eng}},
pages = {{345--359}},
publisher = {{Humana Press}},
series = {{Methods in Molecular Biology}},
title = {{Arrayed molecular barcoding of leukemic stem cells}},
url = {{http://dx.doi.org/10.1007/978-1-0716-0810-4_21}},
doi = {{10.1007/978-1-0716-0810-4_21}},
volume = {{2185}},
year = {{2021}},
}