Cargando…

Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies

A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC) line H1...

Descripción completa

Detalles Bibliográficos
Autores principales: Wu, Youjun, Chang, Tammy, Long, Yan, Huang, He, Kandeel, Fouad, Yee, Jiing-Kuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6909005/
https://www.ncbi.nlm.nih.gov/pubmed/31821946
http://dx.doi.org/10.1016/j.isci.2019.11.038
_version_ 1783478861997015040
author Wu, Youjun
Chang, Tammy
Long, Yan
Huang, He
Kandeel, Fouad
Yee, Jiing-Kuan
author_facet Wu, Youjun
Chang, Tammy
Long, Yan
Huang, He
Kandeel, Fouad
Yee, Jiing-Kuan
author_sort Wu, Youjun
collection PubMed
description A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC) line H1 for specific eradication of undifferentiated cells without affecting differentiated cells. This locus was chosen over NANOG and OCT4, two other well-characterized stem cell loci, due to significantly reduced off-target effect. We showed that undifferentiated H1-iC9 cells were induced to apoptosis by iC9 inducer AP1903, whereas differentiated cell lineages including hematopoietic cells, neurons, and islet beta-like cells were not affected. We also showed that AP1903 selectively removed undifferentiated H1-iC9 cells from a mixed cell population. This strategy therefore provides a layer of safety control before transplantation of a stem-cell-derived product in therapy.
format Online
Article
Text
id pubmed-6909005
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-69090052019-12-23 Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies Wu, Youjun Chang, Tammy Long, Yan Huang, He Kandeel, Fouad Yee, Jiing-Kuan iScience Article A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC) line H1 for specific eradication of undifferentiated cells without affecting differentiated cells. This locus was chosen over NANOG and OCT4, two other well-characterized stem cell loci, due to significantly reduced off-target effect. We showed that undifferentiated H1-iC9 cells were induced to apoptosis by iC9 inducer AP1903, whereas differentiated cell lineages including hematopoietic cells, neurons, and islet beta-like cells were not affected. We also showed that AP1903 selectively removed undifferentiated H1-iC9 cells from a mixed cell population. This strategy therefore provides a layer of safety control before transplantation of a stem-cell-derived product in therapy. Elsevier 2019-11-25 /pmc/articles/PMC6909005/ /pubmed/31821946 http://dx.doi.org/10.1016/j.isci.2019.11.038 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Wu, Youjun
Chang, Tammy
Long, Yan
Huang, He
Kandeel, Fouad
Yee, Jiing-Kuan
Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
title Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
title_full Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
title_fullStr Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
title_full_unstemmed Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
title_short Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
title_sort using gene editing to establish a safeguard system for pluripotent stem-cell-based therapies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6909005/
https://www.ncbi.nlm.nih.gov/pubmed/31821946
http://dx.doi.org/10.1016/j.isci.2019.11.038
work_keys_str_mv AT wuyoujun usinggeneeditingtoestablishasafeguardsystemforpluripotentstemcellbasedtherapies
AT changtammy usinggeneeditingtoestablishasafeguardsystemforpluripotentstemcellbasedtherapies
AT longyan usinggeneeditingtoestablishasafeguardsystemforpluripotentstemcellbasedtherapies
AT huanghe usinggeneeditingtoestablishasafeguardsystemforpluripotentstemcellbasedtherapies
AT kandeelfouad usinggeneeditingtoestablishasafeguardsystemforpluripotentstemcellbasedtherapies
AT yeejiingkuan usinggeneeditingtoestablishasafeguardsystemforpluripotentstemcellbasedtherapies