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High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing

We have reported that of the 10 commonly used AAV serotype vectors, AAV6 is the most efficient in transducing primary human hematopoietic stem/progenitor cells (HSPCs). However, the transduction efficiency of the wild-type (WT) AAV6 vector varies greatly in HSPCs from different donors. Here we repor...

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Autores principales: Ling, Chen, Bhukhai, Kanit, Yin, Zifei, Tan, Mengqun, Yoder, Mervin C., Leboulch, Philippe, Payen, Emmanuel, Srivastava, Arun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5069717/
https://www.ncbi.nlm.nih.gov/pubmed/27759036
http://dx.doi.org/10.1038/srep35495
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author Ling, Chen
Bhukhai, Kanit
Yin, Zifei
Tan, Mengqun
Yoder, Mervin C.
Leboulch, Philippe
Payen, Emmanuel
Srivastava, Arun
author_facet Ling, Chen
Bhukhai, Kanit
Yin, Zifei
Tan, Mengqun
Yoder, Mervin C.
Leboulch, Philippe
Payen, Emmanuel
Srivastava, Arun
author_sort Ling, Chen
collection PubMed
description We have reported that of the 10 commonly used AAV serotype vectors, AAV6 is the most efficient in transducing primary human hematopoietic stem/progenitor cells (HSPCs). However, the transduction efficiency of the wild-type (WT) AAV6 vector varies greatly in HSPCs from different donors. Here we report two distinct strategies to further increase the transduction efficiency in HSPCs from donors that are transduced less efficiently with the WT AAV6 vectors. The first strategy involved modifications of the viral capsid proteins where specific surface-exposed tyrosine (Y) and threonine (T) residues were mutagenized to generate a triple-mutant (Y705 + Y731F + T492V) AAV6 vector. The second strategy involved the use of ex vivo transduction at high cell density. The combined use of these strategies resulted in transduction efficiency exceeding ~90% in HSPCs at significantly reduced vector doses. Our studies have significant implications in the optimal use of capsid-optimized AAV6 vectors in genome editing in HSPCs.
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spelling pubmed-50697172016-10-26 High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing Ling, Chen Bhukhai, Kanit Yin, Zifei Tan, Mengqun Yoder, Mervin C. Leboulch, Philippe Payen, Emmanuel Srivastava, Arun Sci Rep Article We have reported that of the 10 commonly used AAV serotype vectors, AAV6 is the most efficient in transducing primary human hematopoietic stem/progenitor cells (HSPCs). However, the transduction efficiency of the wild-type (WT) AAV6 vector varies greatly in HSPCs from different donors. Here we report two distinct strategies to further increase the transduction efficiency in HSPCs from donors that are transduced less efficiently with the WT AAV6 vectors. The first strategy involved modifications of the viral capsid proteins where specific surface-exposed tyrosine (Y) and threonine (T) residues were mutagenized to generate a triple-mutant (Y705 + Y731F + T492V) AAV6 vector. The second strategy involved the use of ex vivo transduction at high cell density. The combined use of these strategies resulted in transduction efficiency exceeding ~90% in HSPCs at significantly reduced vector doses. Our studies have significant implications in the optimal use of capsid-optimized AAV6 vectors in genome editing in HSPCs. Nature Publishing Group 2016-10-19 /pmc/articles/PMC5069717/ /pubmed/27759036 http://dx.doi.org/10.1038/srep35495 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International 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/4.0/
spellingShingle Article
Ling, Chen
Bhukhai, Kanit
Yin, Zifei
Tan, Mengqun
Yoder, Mervin C.
Leboulch, Philippe
Payen, Emmanuel
Srivastava, Arun
High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing
title High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing
title_full High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing
title_fullStr High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing
title_full_unstemmed High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing
title_short High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for Overcoming Donor-Variation and Implications in Genome Editing
title_sort high-efficiency transduction of primary human hematopoietic stem/progenitor cells by aav6 vectors: strategies for overcoming donor-variation and implications in genome editing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5069717/
https://www.ncbi.nlm.nih.gov/pubmed/27759036
http://dx.doi.org/10.1038/srep35495
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