High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors

Strategies for Overcoming Donor-Variation and Implications in Genome Editing

Chen Ling, Kanit Bhukhai, Zifei Yin, Mengqun Tan, Mervin Yoder, Philippe Leboulch, Emmanuel Payen, Arun Srivastava

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article number35495
JournalScientific Reports
Volume6
DOIs
StatePublished - Oct 19 2016

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Hematopoietic Stem Cells
Gene Editing
Capsid
Capsid Proteins
Viral Proteins
Threonine
Tyrosine
Cell Count

ASJC Scopus subject areas

  • General

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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. / Ling, Chen; Bhukhai, Kanit; Yin, Zifei; Tan, Mengqun; Yoder, Mervin; Leboulch, Philippe; Payen, Emmanuel; Srivastava, Arun.

In: Scientific Reports, Vol. 6, 35495, 19.10.2016.

Research output: Contribution to journalArticle

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abstract = "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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