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Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation

Genome editing with site-specific endonucleases has implications for basic biomedical research as well as for gene therapy. We generated helper-dependent, capsid-modified adenovirus (HD-Ad5/35) vectors for zinc-finger nuclease (ZFN)– or transcription activator-like effector nuclease (TALEN)–mediated...

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Autores principales: Saydaminova, Kamola, Ye, Xun, Wang, Hongjie, Richter, Maximilian, Ho, Martin, Chen, HongZhuan, Xu, Ning, Kim, Jin-Soo, Papapetrou, Eirini, Holmes, Michael C, Gregory, Philip D, Palmer, Donna, Ng, Philip, Ehrhardt, Anja, Lieber, André
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4448996/
https://www.ncbi.nlm.nih.gov/pubmed/26052525
http://dx.doi.org/10.1038/mtm.2014.57
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author Saydaminova, Kamola
Ye, Xun
Wang, Hongjie
Richter, Maximilian
Ho, Martin
Chen, HongZhuan
Xu, Ning
Kim, Jin-Soo
Papapetrou, Eirini
Holmes, Michael C
Gregory, Philip D
Palmer, Donna
Ng, Philip
Ehrhardt, Anja
Lieber, André
author_facet Saydaminova, Kamola
Ye, Xun
Wang, Hongjie
Richter, Maximilian
Ho, Martin
Chen, HongZhuan
Xu, Ning
Kim, Jin-Soo
Papapetrou, Eirini
Holmes, Michael C
Gregory, Philip D
Palmer, Donna
Ng, Philip
Ehrhardt, Anja
Lieber, André
author_sort Saydaminova, Kamola
collection PubMed
description Genome editing with site-specific endonucleases has implications for basic biomedical research as well as for gene therapy. We generated helper-dependent, capsid-modified adenovirus (HD-Ad5/35) vectors for zinc-finger nuclease (ZFN)– or transcription activator-like effector nuclease (TALEN)–mediated genome editing in human CD34+ hematopoietic stem cells (HSCs) from mobilized adult donors. The production of these vectors required that ZFN and TALEN expression in HD-Ad5/35 producer 293-Cre cells was suppressed. To do this, we developed a microRNA (miRNA)-based system for regulation of gene expression based on miRNA expression profiling of 293-Cre and CD34+ cells. Using miR-183-5p and miR-218-5p based regulation of transgene gene expression, we first produced an HD-Ad5/35 vector expressing a ZFN specific to the HIV coreceptor gene ccr5. We demonstrated that HD-Ad5/35.ZFNmiR vector conferred ccr5 knock out in primitive HSC (i.e., long-term culture initiating cells and NOD/SCID repopulating cells). The ccr5 gene disruption frequency achieved in engrafted HSCs found in the bone marrow of transplanted mice is clinically relevant for HIV therapy considering that these cells can give rise to multiple lineages, including all the lineages that represent targets and reservoirs for HIV. We produced a second HD-Ad5/35 vector expressing a TALEN targeting the DNase hypersensitivity region 2 (HS2) within the globin locus control region. This vector has potential for targeted gene correction in hemoglobinopathies. The miRNA regulated HD-Ad5/35 vector platform for expression of site-specific endonucleases has numerous advantages over currently used vectors as a tool for genome engineering of HSCs for therapeutic purposes.
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spelling pubmed-44489962015-06-05 Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation Saydaminova, Kamola Ye, Xun Wang, Hongjie Richter, Maximilian Ho, Martin Chen, HongZhuan Xu, Ning Kim, Jin-Soo Papapetrou, Eirini Holmes, Michael C Gregory, Philip D Palmer, Donna Ng, Philip Ehrhardt, Anja Lieber, André Mol Ther Methods Clin Dev Article Genome editing with site-specific endonucleases has implications for basic biomedical research as well as for gene therapy. We generated helper-dependent, capsid-modified adenovirus (HD-Ad5/35) vectors for zinc-finger nuclease (ZFN)– or transcription activator-like effector nuclease (TALEN)–mediated genome editing in human CD34+ hematopoietic stem cells (HSCs) from mobilized adult donors. The production of these vectors required that ZFN and TALEN expression in HD-Ad5/35 producer 293-Cre cells was suppressed. To do this, we developed a microRNA (miRNA)-based system for regulation of gene expression based on miRNA expression profiling of 293-Cre and CD34+ cells. Using miR-183-5p and miR-218-5p based regulation of transgene gene expression, we first produced an HD-Ad5/35 vector expressing a ZFN specific to the HIV coreceptor gene ccr5. We demonstrated that HD-Ad5/35.ZFNmiR vector conferred ccr5 knock out in primitive HSC (i.e., long-term culture initiating cells and NOD/SCID repopulating cells). The ccr5 gene disruption frequency achieved in engrafted HSCs found in the bone marrow of transplanted mice is clinically relevant for HIV therapy considering that these cells can give rise to multiple lineages, including all the lineages that represent targets and reservoirs for HIV. We produced a second HD-Ad5/35 vector expressing a TALEN targeting the DNase hypersensitivity region 2 (HS2) within the globin locus control region. This vector has potential for targeted gene correction in hemoglobinopathies. The miRNA regulated HD-Ad5/35 vector platform for expression of site-specific endonucleases has numerous advantages over currently used vectors as a tool for genome engineering of HSCs for therapeutic purposes. Nature Publishing Group 2015-01-14 /pmc/articles/PMC4448996/ /pubmed/26052525 http://dx.doi.org/10.1038/mtm.2014.57 Text en Copyright © 2015 American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed. under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
spellingShingle Article
Saydaminova, Kamola
Ye, Xun
Wang, Hongjie
Richter, Maximilian
Ho, Martin
Chen, HongZhuan
Xu, Ning
Kim, Jin-Soo
Papapetrou, Eirini
Holmes, Michael C
Gregory, Philip D
Palmer, Donna
Ng, Philip
Ehrhardt, Anja
Lieber, André
Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation
title Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation
title_full Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation
title_fullStr Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation
title_full_unstemmed Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation
title_short Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation
title_sort efficient genome editing in hematopoietic stem cells with helper-dependent ad5/35 vectors expressing site-specific endonucleases under microrna regulation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4448996/
https://www.ncbi.nlm.nih.gov/pubmed/26052525
http://dx.doi.org/10.1038/mtm.2014.57
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