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Short DNA Hairpins Compromise Recombinant Adeno-Associated Virus Genome Homogeneity.
Xie, Jun; Mao, Qin; Tai, Phillip W L; He, Ran; Ai, Jianzhong; Su, Qin; Zhu, Ye; Ma, Hong; Li, Jia; Gong, Shoufang; Wang, Dan; Gao, Zhen; Li, Mengxin; Zhong, Li; Zhou, Heather; Gao, Guangping.
Affiliation
  • Xie J; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01
  • Mao Q; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Thoracic Cancer, Cancer Center, West China Hospital, West China School of Clinical Medicine, Sichuan University, Chengdu, Sichuan 610000, China.
  • Tai PWL; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • He R; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Ai J; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Urology Department, West China Hospital, Sichuan University, Chengdu, Sichuan 610000, China.
  • Su Q; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Zhu Y; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Cardiovascular Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan 610000, China.
  • Ma H; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Li J; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Gong S; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Wang D; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Gao Z; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Li M; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Zhong L; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • Zhou H; Genetics and Pharmacogenomics, Merck Research Laboratory, Kenilworth, NJ 07033, USA. Electronic address: hui_zhou@merck.com.
  • Gao G; Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA; Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01
Mol Ther ; 25(6): 1363-1374, 2017 06 07.
Article in En | MEDLINE | ID: mdl-28462820
ABSTRACT
Short hairpin (sh)RNAs delivered by recombinant adeno-associated viruses (rAAVs) are valuable tools to study gene function in vivo and a promising gene therapy platform. Our data show that incorporation of shRNA transgenes into rAAV constructs reduces vector yield and produces a population of truncated and defective genomes. We demonstrate that sequences with hairpins or hairpin-like structures drive the generation of truncated AAV genomes through a polymerase redirection mechanism during viral genome replication. Our findings reveal the importance of genomic secondary structure when optimizing viral vector designs. We also discovered that shDNAs could be adapted to act as surrogate mutant inverted terminal repeats (mTRs), sequences that were previously thought to be required for functional self-complementary AAV vectors. The use of shDNAs as artificial mTRs opens the door to engineering a new generation of AAV vectors with improved potency, genetic stability, and safety for both preclinical studies and human gene therapy.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: DNA, Viral / Genome, Viral / Dependovirus / Inverted Repeat Sequences / Genetic Vectors Type of study: Prognostic_studies / Risk_factors_studies Limits: Animals / Humans / Male Language: En Journal: Mol Ther Journal subject: BIOLOGIA MOLECULAR / TERAPEUTICA Year: 2017 Document type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: DNA, Viral / Genome, Viral / Dependovirus / Inverted Repeat Sequences / Genetic Vectors Type of study: Prognostic_studies / Risk_factors_studies Limits: Animals / Humans / Male Language: En Journal: Mol Ther Journal subject: BIOLOGIA MOLECULAR / TERAPEUTICA Year: 2017 Document type: Article