A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.

A major challenge in the stem cell biology field is the ability to produce fully functional cells from induced pluripotent stem cells (iPSCs) that are a valuable resource for cell therapy, drug screening, and disease modelling. Here, we developed a novel inducible CRISPR-mediated activation strategy...

Full description

Bibliographic Details
Authors: Petazzi, P, Ventura, T, Luongo, FP, McClafferty, H, May, A, Taylor, HA, Shipston, MJ, Romanò, N, Forrester, LM, Menendez, P, Fidanza, A
Format: article
Status:Published version
Publication Date:2024
Country:España
Institution:Fundació Sant Joan de Déu
Repository:r-FSJD. Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déu
OAI Identifier:oai:dnet:r-fsjd______::748eb784c5331eba6b485b369f25a3fc
Online Access:https://fsjd.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=28271
Access Level:Open access
Keyword:CRISPR activation
developmental biology
haematopoiesis
human
pluripotent stem cells
regenerative medicine
stem cells
id ES_c3efab7cc9cdbf6ebce9ef75fe57281a
oai_identifier_str oai:dnet:r-fsjd______::748eb784c5331eba6b485b369f25a3fc
network_acronym_str ES
network_name_str España
repository_id_str
spelling A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.Petazzi, PVentura, TLuongo, FPMcClafferty, HMay, ATaylor, HAShipston, MJRomanò, NForrester, LMMenendez, PFidanza, ACRISPR activationdevelopmental biologyhaematopoiesishumanpluripotent stem cellsregenerative medicinestem cellsA major challenge in the stem cell biology field is the ability to produce fully functional cells from induced pluripotent stem cells (iPSCs) that are a valuable resource for cell therapy, drug screening, and disease modelling. Here, we developed a novel inducible CRISPR-mediated activation strategy (iCRISPRa) to drive the expression of multiple endogenous transcription factors (TFs) important for in vitro cell fate and differentiation of iPSCs to haematopoietic progenitor cells. This work has identified a key role for IGFBP2 in developing haematopoietic progenitors. We first identified nine candidate TFs that we predicted to be involved in blood cell emergence during development, then generated tagged gRNAs directed to the transcriptional start site of these TFs that could also be detected during single-cell RNA sequencing (scRNAseq). iCRISPRa activation of these endogenous TFs resulted in a significant expansion of arterial-fated endothelial cells expressing high levels of IGFBP2, and our analysis indicated that IGFBP2 is involved in the remodelling of metabolic activity during in vitro endothelial to haematopoietic transition. As well as providing fundamental new insights into the mechanisms of haematopoietic differentiation, the broader applicability of iCRISPRa provides a valuable tool for studying dynamic processes in development and for recapitulating abnormal phenotypes characterised by ectopic activation of specific endogenous gene expression in a wide range of systems.eLIFE SCIENCES PUBL LTD2024info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://fsjd.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=28271eLifeISSN: 2050084Xreponame:r-FSJD. Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déuinstname:Fundació Sant Joan de DéuInglésinfo:eu-repo/semantics/openAccessoai:dnet:r-fsjd______::748eb784c5331eba6b485b369f25a3fc2026-05-27T12:37:41Z
dc.title.none.fl_str_mv A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
title A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
spellingShingle A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
Petazzi, P
CRISPR activation
developmental biology
haematopoiesis
human
pluripotent stem cells
regenerative medicine
stem cells
title_short A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
title_full A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
title_fullStr A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
title_full_unstemmed A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
title_sort A novel human pluripotent stem cell gene activation system identifies IGFBP2 as a mediator in the production of haematopoietic progenitors in vitro.
dc.creator.none.fl_str_mv Petazzi, P
Ventura, T
Luongo, FP
McClafferty, H
May, A
Taylor, HA
Shipston, MJ
Romanò, N
Forrester, LM
Menendez, P
Fidanza, A
author Petazzi, P
author_facet Petazzi, P
Ventura, T
Luongo, FP
McClafferty, H
May, A
Taylor, HA
Shipston, MJ
Romanò, N
Forrester, LM
Menendez, P
Fidanza, A
author_role author
author2 Ventura, T
Luongo, FP
McClafferty, H
May, A
Taylor, HA
Shipston, MJ
Romanò, N
Forrester, LM
Menendez, P
Fidanza, A
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CRISPR activation
developmental biology
haematopoiesis
human
pluripotent stem cells
regenerative medicine
stem cells
topic CRISPR activation
developmental biology
haematopoiesis
human
pluripotent stem cells
regenerative medicine
stem cells
description A major challenge in the stem cell biology field is the ability to produce fully functional cells from induced pluripotent stem cells (iPSCs) that are a valuable resource for cell therapy, drug screening, and disease modelling. Here, we developed a novel inducible CRISPR-mediated activation strategy (iCRISPRa) to drive the expression of multiple endogenous transcription factors (TFs) important for in vitro cell fate and differentiation of iPSCs to haematopoietic progenitor cells. This work has identified a key role for IGFBP2 in developing haematopoietic progenitors. We first identified nine candidate TFs that we predicted to be involved in blood cell emergence during development, then generated tagged gRNAs directed to the transcriptional start site of these TFs that could also be detected during single-cell RNA sequencing (scRNAseq). iCRISPRa activation of these endogenous TFs resulted in a significant expansion of arterial-fated endothelial cells expressing high levels of IGFBP2, and our analysis indicated that IGFBP2 is involved in the remodelling of metabolic activity during in vitro endothelial to haematopoietic transition. As well as providing fundamental new insights into the mechanisms of haematopoietic differentiation, the broader applicability of iCRISPRa provides a valuable tool for studying dynamic processes in development and for recapitulating abnormal phenotypes characterised by ectopic activation of specific endogenous gene expression in a wide range of systems.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://fsjd.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=28271
url https://fsjd.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=28271
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv eLIFE SCIENCES PUBL LTD
publisher.none.fl_str_mv eLIFE SCIENCES PUBL LTD
dc.source.none.fl_str_mv eLife
ISSN: 2050084X
reponame:r-FSJD. Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déu
instname:Fundació Sant Joan de Déu
instname_str Fundació Sant Joan de Déu
reponame_str r-FSJD. Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déu
collection r-FSJD. Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déu
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869418838480650240
score 15,81155

Similar Items