Structure and function of the adeno-associated virus (AAV) genome will be studied at the molecular level. Use will be made of a recombinant plasmid in which the AAV genome is inserted into pBR322. When this plasmid is transfected into human cells with helper, the viral genome is rescued and replicated. With this system alterations can be made at any place in the AAV genome and the effects on replication directly assessed. Problems to be studied include: 1) structural and sequence requirements in the inverted terminal repetition for DNA replication and rescue of the integrated genome, 2) identification of AAV sequences that regulate transcription, 3) identification of helper functions required for AAV transcription, 4) identification of additional spliced species of AAV transcripts, and 5) identification of nonstructural proteins. Additional projects include isolation of nonstructural proteins, and characterization of AAV sequences integrated into cellular DNA and in AAV-SV40 recombinants by cloning, restriction analysis, and sequence determination. Finally, computer analysis will be used to search for interesting sequence arrangements and potential secondary structures that might affect regulation of transcription and translation. Sequences that are so identified will be modified and the effect on transcription and translation determined using the biologically active clone described above.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI022251-02
Application #
3133137
Study Section
Virology Study Section (VR)
Project Start
1984-08-02
Project End
1989-07-31
Budget Start
1985-08-01
Budget End
1986-07-31
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Weill Medical College of Cornell University
Department
Type
Schools of Medicine
DUNS #
201373169
City
New York
State
NY
Country
United States
Zip Code
10065
Berns, K I; Meneses, P; Duvoisin, R et al. (2001) Use of recombinant angiostatin to prevent retinal neovascularization. Trans Am Clin Climatol Assoc 112:68-75; discussion 75-6
Meneses, P I; Hajjar, K A; Berns, K I et al. (2001) Recombinant angiostatin prevents retinal neovascularization in a murine proliferative retinopathy model. Gene Ther 8:646-8
Dyall, J; Szabo, P; Berns, K I (1999) Adeno-associated virus (AAV) site-specific integration: formation of AAV-AAVS1 junctions in an in vitro system. Proc Natl Acad Sci U S A 96:12849-54
Linden, R M; Winocour, E; Berns, K I (1996) The recombination signals for adeno-associated virus site-specific integration. Proc Natl Acad Sci U S A 93:7966-72
Linden, R M; Ward, P; Giraud, C et al. (1996) Site-specific integration by adeno-associated virus. Proc Natl Acad Sci U S A 93:11288-94
Berns, K I; Linden, R M (1995) The cryptic life style of adeno-associated virus. Bioessays 17:237-45
Leonard, C J; Berns, K I (1994) Cloning, expression, and partial purification of Rep78: an adeno-associated virus replication protein. Virology 200:566-73
Leonard, C J; Berns, K I (1994) Adeno-associated virus type 2: a latent life cycle. Prog Nucleic Acid Res Mol Biol 48:29-52
Giraud, C; Winocour, E; Berns, K I (1994) Site-specific integration by adeno-associated virus is directed by a cellular DNA sequence. Proc Natl Acad Sci U S A 91:10039-43
Hong, G; Ward, P; Berns, K I (1992) In vitro replication of adeno-associated virus DNA. Proc Natl Acad Sci U S A 89:4673-7

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