scholarly journals Adeno-Associated Virus Type 2 Rep40 Modulates the Proliferation Rate of Rep52-Expressing HeLa Cells

Intervirology ◽  
2003 ◽  
Vol 46 (2) ◽  
pp. 127-134
Author(s):  
Donglan Li ◽  
Saburo Kashii ◽  
Toshiyuki Sahara ◽  
Kazuhiko Ito
Keyword(s):  
Virus Type ◽  
Hela Cells ◽  
Proliferation Rate ◽  
Adeno Associated Virus

scholarly journals Rescue and Autonomous Replication of Adeno-Associated Virus Type 2 Genomes Containing Rep-Binding Site Mutations in the Viral p5 Promoter

1998 ◽  
Vol 72 (6) ◽  
pp. 4811-4818 ◽  
Author(s):  
Xu-Shan Wang ◽  
Arun Srivastava
Keyword(s):  
Binding Site ◽  
Virus Type ◽  
Hela Cells ◽  
Biologically Active ◽  
Viral Gene ◽  
Adeno Associated Virus ◽  
Autonomous Replication ◽  
293 Cells ◽  
Rep Proteins

ABSTRACT The Rep proteins encoded by the adeno-associated virus type 2 (AAV) play a crucial role in the rescue, replication, and integration of the viral genome. In the absence of a helper virus, little expression of the AAV Rep proteins occurs, and the AAV genome fails to undergo DNA replication. Since previous studies have established that expression of the Rep78 and Rep68 proteins from the viral p5 promoter is controlled by the Rep-binding site (RBS) and the YY1 factor-binding site (YBS), we constructed a number of recombinant AAV plasmids containing mutations and/or deletions of the RBS and the YBS in the p5 promoter. These plasmids were transfected in HeLa or 293 cells and analyzed for the potential to undergo AAV DNA rescue and replication. Our studies revealed that (i) a low-level rescue and autonomous replication of the wild-type AAV genome occurred in 293 but not in HeLa cells; (ii) mutations in the RBS resulted in augmented expression from the p5 promoter, leading to more efficient rescue and/or replication of the AAV genome in 293 but not in HeLa cells; (iii) little rescue and/or replication occurred from plasmids containing mutations in the YBS alone in the absence of coinfection with adenovirus; (iv) expression of the adenovirus E1A gene products was insufficient to mediate rescue and/or replication of the AAV genome in HeLa cells; (v) autonomously replicated AAV genomes in 293 cells were successfully encapsidated in mature progeny virions that were biologically active in secondary infection of HeLa cells in the presence of adenovirus; and (vi) stable transfection of recombinant AAV plasmids containing a gene for resistance to neomycin significantly affected stable integration only in 293 cells, presumably because rescue and autonomous replication of the AAV genome from these plasmids occurred in 293 cells but not in HeLa or KB cells. These data suggest that in the absence of adenovirus, the AAV Rep protein-RBS interaction plays a dominant role in down-regulating viral gene expression from the p5 promoter and that perturbation in this interaction is sufficient to confer autonomous replication competence to AAV in 293 cells.

Biologically active Rep proteins of adeno-associated virus type 2 produced as fusion proteins in Escherichia coli.

1994 ◽  
Vol 68 (2) ◽  
pp. 797-804 ◽  
Author(s):  
J A Chiorini ◽  
M D Weitzman ◽  
R A Owens ◽  
E Urcelay ◽  
B Safer ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virus Type ◽  
Fusion Proteins ◽  
Biologically Active ◽  
Adeno Associated Virus ◽  
Rep Proteins

scholarly journals Biodistribution of adeno‐associated virus type 2 carrying multi‐characteristic opsin in dogs following intravitreal injection

2021 ◽  
Vol 25 (18) ◽  
pp. 8676-8686
Author(s):  
Kissaou T. Tchedre ◽  
Subrata Batabyal ◽  
Melissa Galicia ◽  
Darryl Narcisse ◽  
Sourajit Mitra Mustafi ◽  
...  
Keyword(s):  
Intravitreal Injection ◽  
Virus Type ◽  
Adeno Associated Virus

scholarly journals Adeno-Associated Virus Type 2 and Hepatocellular Carcinoma?

2015 ◽  
Vol 26 (12) ◽  
pp. 779-781 ◽  
Author(s):  
Kenneth I. Berns ◽  
Barry J. Byrne ◽  
Terence R. Flotte ◽  
Guangping Gao ◽  
William W. Hauswirth ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Virus Type ◽  
Adeno Associated Virus

Induction of an Antitumoral Immune Response by Wild-Type Adeno-Associated Virus Type 2 in an In Vivo Model of Pancreatic Carcinoma

Pancreas ◽  
2007 ◽  
Vol 35 (1) ◽  
pp. 63-72 ◽  
Author(s):  
Sven Eisold ◽  
Jan Schmidt ◽  
Eduard Ryschich ◽  
Michael Gock ◽  
Ernst Klar ◽  
...  
Keyword(s):  
Immune Response ◽  
Pancreatic Carcinoma ◽  
Virus Type ◽  
In Vivo Model ◽  
Wild Type ◽  
Adeno Associated Virus

scholarly journals Presence of atrs-Like Motif Promotes Rep-Mediated Wild-Type Adeno-Associated Virus Type 2 Integration

2015 ◽  
Vol 89 (14) ◽  
pp. 7428-7432 ◽  
Author(s):  
Karl Petri ◽  
Richard Gabriel ◽  
Leticia Agundez ◽  
Raffaele Fronza ◽  
Saira Afzal ◽  
...  
Keyword(s):  
Virus Type ◽  
Integration Site ◽  
Dermal Fibroblasts ◽  
Wild Type ◽  
Mobility Shift ◽  
Adeno Associated Virus ◽  
Rep Protein ◽  
Electrophoretic Mobility Shift Assays ◽  
First Time

High-throughput integration site (IS) analysis of wild-type adeno-associated virus type 2 (wtAAV2) in human dermal fibroblasts (HDFs) and HeLa cells revealed that juxtaposition of a Rep binding site (RBS) and terminal resolution site (trs)-like motif leads to a 4-fold-increased probability of wtAAV integration. Electrophoretic mobility shift assays (EMSAs) confirmed binding of Rep to off-target RBSs. For the first time, we show Rep protein off-target nicking activity, highlighting the importance of the nicking substrate for Rep-mediated integration.

scholarly journals Charge-to-Alanine Mutagenesis of the Adeno-Associated Virus Type 2 Rep78/68 Proteins Yields Temperature-Sensitive and Magnesium-Dependent Variants

2000 ◽  
Vol 74 (1) ◽  
pp. 591-591
Author(s):  
Denise K. Gavin ◽  
Samuel M. Young ◽  
Weidong Xiao ◽  
Brenda Temple ◽  
Corinne R. Abernathy ◽  
...  
Keyword(s):  
Virus Type ◽  
Temperature Sensitive ◽  
Adeno Associated Virus

scholarly journals Genetic modifications of the adeno-associated virus type 2 capsid reduce the affinity and the neutralizing effects of human serum antibodies

Gene Therapy ◽  
2003 ◽  
Vol 10 (26) ◽  
pp. 2139-2147 ◽  
Author(s):  
N A Huttner ◽  
A Girod ◽  
L Perabo ◽  
D Edbauer ◽  
J A Kleinschmidt ◽  
...  
Keyword(s):  
Human Serum ◽  
Virus Type ◽  
Serum Antibodies ◽  
Adeno Associated Virus ◽  
Genetic Modifications
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