Recombinant adeno-associated virus vector for gene transfer to the transplanted rat heart
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Recombinant adeno-associated virus vector for gene transfer to the transplanted rat heart. / Schirmer, Johannes M; Miyagi, Naoto; Rao, Vinay P; Ricci, Davide; Federspiel, Mark J; Kotin, Robert M; Russell, Stephen J; McGregor, Christopher G A.
in: TRANSPL INT, Jahrgang 20, Nr. 6, 06.2007, S. 550-557.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Recombinant adeno-associated virus vector for gene transfer to the transplanted rat heart
AU - Schirmer, Johannes M
AU - Miyagi, Naoto
AU - Rao, Vinay P
AU - Ricci, Davide
AU - Federspiel, Mark J
AU - Kotin, Robert M
AU - Russell, Stephen J
AU - McGregor, Christopher G A
PY - 2007/6
Y1 - 2007/6
N2 - Efficient durable viral vector transduction of the transplanted heart remains elusive. This study assesses the potential of recombinant adeno-associated virus (rAAV) mediated gene delivery to the transplanted rat heart. rAAV serotype 1, 2 and 5 vectors encoding the green fluorescent protein (GFP) gene (1 x 10(11) viral particles/ml) were diluted in cold University of Wisconsin solution and circulated through the coronary vasculature of the donor organs for 30 min before syngeneic rat heterotopic heart transplantation was performed. Study 1: animals (n = 5 each serotype) were killed at 21 days post-transplant to evaluate the efficiency of GFP transduction using RT-PCR and expression by fluorescence microscopy. Study 2: using rAAV-1, animals (n = 5 each group) were killed at 7, 21 and 84 days to evaluate the durability of GFP expression. The maximum cardiac GFP expression at 21 days was observed in rAAV-1. GFP expression by rAAV-1 was detectable at 7 days, improved at 21 days, and was still evident at 84 days. This study demonstrates cardiac rAAV gene transduction with a cold perfusion preservation system of the donor heart. These data show that AAV-1 is superior to AAV-2 and AAV-5 for this purpose and that durable expression is achievable.
AB - Efficient durable viral vector transduction of the transplanted heart remains elusive. This study assesses the potential of recombinant adeno-associated virus (rAAV) mediated gene delivery to the transplanted rat heart. rAAV serotype 1, 2 and 5 vectors encoding the green fluorescent protein (GFP) gene (1 x 10(11) viral particles/ml) were diluted in cold University of Wisconsin solution and circulated through the coronary vasculature of the donor organs for 30 min before syngeneic rat heterotopic heart transplantation was performed. Study 1: animals (n = 5 each serotype) were killed at 21 days post-transplant to evaluate the efficiency of GFP transduction using RT-PCR and expression by fluorescence microscopy. Study 2: using rAAV-1, animals (n = 5 each group) were killed at 7, 21 and 84 days to evaluate the durability of GFP expression. The maximum cardiac GFP expression at 21 days was observed in rAAV-1. GFP expression by rAAV-1 was detectable at 7 days, improved at 21 days, and was still evident at 84 days. This study demonstrates cardiac rAAV gene transduction with a cold perfusion preservation system of the donor heart. These data show that AAV-1 is superior to AAV-2 and AAV-5 for this purpose and that durable expression is achievable.
KW - Adenoviridae
KW - Animals
KW - Gene Expression
KW - Genetic Vectors/administration & dosage
KW - Green Fluorescent Proteins
KW - Heart Transplantation
KW - Male
KW - Rats
KW - Rats, Inbred Lew
KW - Recombination, Genetic
KW - Reverse Transcriptase Polymerase Chain Reaction
KW - Transduction, Genetic/methods
U2 - 10.1111/j.1432-2277.2007.00479.x
DO - 10.1111/j.1432-2277.2007.00479.x
M3 - SCORING: Journal article
C2 - 17403107
VL - 20
SP - 550
EP - 557
JO - TRANSPL INT
JF - TRANSPL INT
SN - 0934-0874
IS - 6
ER -
