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Fast but not furious: a streamlined selection method for genome-edited cells

In the last decade, transcription activator-like effector nucleases and CRISPR-based genome engineering have revolutionized our approach to biology. Because of their high efficiency and ease of use, the development of custom knock-out and knock-in animal or cell models is now within reach for almost...

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Autores principales: Ramachandran, Haribaskar, Martins, Soraia, Kontarakis, Zacharias, Krutmann, Jean, Rossi, Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Life Science Alliance LLC 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8127327/
https://www.ncbi.nlm.nih.gov/pubmed/33903218
http://dx.doi.org/10.26508/lsa.202101051
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author Ramachandran, Haribaskar
Martins, Soraia
Kontarakis, Zacharias
Krutmann, Jean
Rossi, Andrea
author_facet Ramachandran, Haribaskar
Martins, Soraia
Kontarakis, Zacharias
Krutmann, Jean
Rossi, Andrea
author_sort Ramachandran, Haribaskar
collection PubMed
description In the last decade, transcription activator-like effector nucleases and CRISPR-based genome engineering have revolutionized our approach to biology. Because of their high efficiency and ease of use, the development of custom knock-out and knock-in animal or cell models is now within reach for almost every laboratory. Nonetheless, the generation of genetically modified cells often requires a selection step, usually achieved by antibiotics or fluorescent markers. The choice of the selection marker is based on the available laboratory resources, such as cell types, and parameters such as time and cost should also be taken into consideration. Here, we present a new and fast strategy called magnetic-activated genome-edited cell sorting, to select genetically modified cells based on the ability to magnetically sort surface antigens (i.e., tCD19) present in Cas9-positive cells. By using magnetic-activated genome-edited cell sorting, we successfully generated and isolated genetically modified human-induced pluripotent stem cells, primary human fibroblasts, SH-SY5Y neuroblast-like cells, HaCaT and HEK 293T cells. Our strategy expands the genome editing toolbox by offering a fast, cheap, and an easy to use alternative to the available selection methods.
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spelling pubmed-81273272021-05-21 Fast but not furious: a streamlined selection method for genome-edited cells Ramachandran, Haribaskar Martins, Soraia Kontarakis, Zacharias Krutmann, Jean Rossi, Andrea Life Sci Alliance Research Articles In the last decade, transcription activator-like effector nucleases and CRISPR-based genome engineering have revolutionized our approach to biology. Because of their high efficiency and ease of use, the development of custom knock-out and knock-in animal or cell models is now within reach for almost every laboratory. Nonetheless, the generation of genetically modified cells often requires a selection step, usually achieved by antibiotics or fluorescent markers. The choice of the selection marker is based on the available laboratory resources, such as cell types, and parameters such as time and cost should also be taken into consideration. Here, we present a new and fast strategy called magnetic-activated genome-edited cell sorting, to select genetically modified cells based on the ability to magnetically sort surface antigens (i.e., tCD19) present in Cas9-positive cells. By using magnetic-activated genome-edited cell sorting, we successfully generated and isolated genetically modified human-induced pluripotent stem cells, primary human fibroblasts, SH-SY5Y neuroblast-like cells, HaCaT and HEK 293T cells. Our strategy expands the genome editing toolbox by offering a fast, cheap, and an easy to use alternative to the available selection methods. Life Science Alliance LLC 2021-04-26 /pmc/articles/PMC8127327/ /pubmed/33903218 http://dx.doi.org/10.26508/lsa.202101051 Text en © 2021 Ramachandran et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Articles
Ramachandran, Haribaskar
Martins, Soraia
Kontarakis, Zacharias
Krutmann, Jean
Rossi, Andrea
Fast but not furious: a streamlined selection method for genome-edited cells
title Fast but not furious: a streamlined selection method for genome-edited cells
title_full Fast but not furious: a streamlined selection method for genome-edited cells
title_fullStr Fast but not furious: a streamlined selection method for genome-edited cells
title_full_unstemmed Fast but not furious: a streamlined selection method for genome-edited cells
title_short Fast but not furious: a streamlined selection method for genome-edited cells
title_sort fast but not furious: a streamlined selection method for genome-edited cells
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8127327/
https://www.ncbi.nlm.nih.gov/pubmed/33903218
http://dx.doi.org/10.26508/lsa.202101051
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