PMID- 27910043
OWN - NLM
STAT- MEDLINE
DCOM- 20180115
LR  - 20181113
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Print)
IS  - 1064-3745 (Linking)
VI  - 1521
DP  - 2017
TI  - Production and Characterization of Vectors Based on the Cardiotropic AAV Serotype 
      9.
PG  - 91-107
AB  - Vectors based on adeno-associated virus serotype 9 (AAV9) efficiently transduce 
      cardiomyocytes in both rodents and large animal models upon either systemic or 
      regional vector delivery. In this chapter, we describe the most widely used 
      production and purification method of AAV9. This production approach does not 
      depend on the use of a helpervirus but instead on transient transfection of 
      HEK293T cells with a plasmid containing the recombinant AAV genome and a second 
      plasmid encoding the AAV9 capsid proteins, the AAV Rep proteins and the 
      adenoviral helper functions. The recombinant AAV is then purified by iodixanol 
      density gradient centrifugation. This chapter also describes in detail the 
      characterization and quality control methods required for assuring high quality 
      vector preparations, which is of particular importance for experiments in large 
      animal models.
FAU - Kohlbrenner, Erik
AU  - Kohlbrenner E
AD  - Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, One 
      Gustave L. Levy Place, Box 1030, New York, NY, 10029-6574, USA.
FAU - Weber, Thomas
AU  - Weber T
AD  - Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, One 
      Gustave L. Levy Place, Box 1030, New York, NY, 10029-6574, USA. 
      thomas.weber@mssm.edu.
LA  - eng
GR  - R01 HL131404/HL/NHLBI NIH HHS/United States
GR  - P50 HL112324/HL/NHLBI NIH HHS/United States
GR  - R01 HL119046/HL/NHLBI NIH HHS/United States
GR  - R01 HL128099/HL/NHLBI NIH HHS/United States
GR  - R01 HL117505/HL/NHLBI NIH HHS/United States
GR  - R01 HL129814/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
SB  - IM
MH  - Capsid/metabolism
MH  - Cloning, Molecular
MH  - Dependovirus/*genetics/ultrastructure
MH  - Dialysis
MH  - Genetic Vectors/*metabolism
MH  - Genome, Viral
MH  - HEK293 Cells
MH  - Humans
MH  - Plasmids/metabolism
MH  - Real-Time Polymerase Chain Reaction
MH  - *Serogroup
MH  - Transfection
MH  - Ultracentrifugation
MH  - Virion/metabolism/ultrastructure
PMC - PMC5863534
MID - NIHMS949928
OTO - NOTNLM
OT  - AAV
OT  - AAV characterization
OT  - AAV production
OT  - AAV9
OT  - Adeno-associated virus
OT  - Cardiotropic
OT  - Gene therapy
OT  - Serotype
EDAT- 2016/12/03 06:00
MHDA- 2018/01/16 06:00
PMCR- 2018/03/22
CRDT- 2016/12/03 06:00
PHST- 2016/12/03 06:00 [entrez]
PHST- 2016/12/03 06:00 [pubmed]
PHST- 2018/01/16 06:00 [medline]
PHST- 2018/03/22 00:00 [pmc-release]
AID - 10.1007/978-1-4939-6588-5_6 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2017;1521:91-107. doi: 10.1007/978-1-4939-6588-5_6.