Nicholas Muzyczka

Long-term consequences of human alpha-synuclein overexpression in the primate ventral midbrain (2007)

Eslamboli, Andisheh, Romero-Ramos, Marina, Burger, Corinna, Bjorklund, Tomas, Muzyczka, Nicholas, Mandel, Ronald J., ...

Overexpression of human α-synuclein (α-syn) using recombinant adeno-associated viral (rAAV) vectors provides a novel tool to study neurodegenerative processes seen in Parkinson's disease and other...

Long-term consequences of human alpha-synuclein overexpression in the primate ventral midbrain (2007)

Eslamboli, Andisheh, Romero-Ramos, Marina, Burger, Corinna, Bjorklund, Tomas, Muzyczka, Nicholas, Mandel, Ronald J., ...

Overexpression of human α-synuclein (α-syn) using recombinant adeno-associated viral (rAAV) vectors provides a novel tool to study neurodegenerative processes seen in Parkinson's disease and other...

Reversal of dyskinesias in an animal model of Parkinson's disease by continuous L-DOPA delivery using rAAV vectors (2005)

Carlsson, Thomas, Winkler, Christian, Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J., Cenci, Angela, ...

Dyskinesias are a major complication of long-term l-3,4-dihydroxyphenylalanine (l-DOPA) treatment in Parkinson's disease, and are believed to result from the intermittent and pulsatile supply of...

Reversal of dyskinesias in an animal model of Parkinson's disease by continuous L-DOPA delivery using rAAV vectors (2005)

Carlsson, Thomas, Winkler, Christian, Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J., Cenci, Angela, ...

Summary Dyskinesias are a major complication of long-term L-3,4-dihydroxyphenylalanine (L-DOPA) treatment in Parkinson's disease, and are believed to result from the intermittent and pulsatile supply...

Reversal of dyskinesias in an animal model of Parkinson's disease by continuous L-DOPA delivery using rAAV vectors (2005)

Carlsson, Thomas, Winkler, Christian, Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J., Cenci, Angela, ...

Summary Dyskinesias are a major complication of long-term L-3,4-dihydroxyphenylalanine (L-DOPA) treatment in Parkinson's disease, and are believed to result from the intermittent and pulsatile supply...

Neuropathological and behavioral consequences of adeno-associated viral vector-mediated continuous intrastriatal neurotrophin delivery in a focal ischemia model in rats. (2002)

Andsberg, Gunnar, Kokaia, Zaal, Klein, Ronald L, Muzyczka, Nicholas, Lindvall, Olle, Mandel, Ronald J

Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) were continuously delivered to the striatum at biologically active levels via recombinant adeno-associated viral (rAAV) gene...

Parkinson-like neurodegeneration induced by targeted overexpression of alpha-synuclein in the nigrostriatal system. (2002)

Kirik, Deniz, Rosenblad, Carl, Burger, Corinna, Lundberg, Cecilia, Johansen, Teit E, Muzyczka, Nicholas, ...

Recombinant adeno-associated viral vectors display efficient tropism for transduction of the dopamine neurons of the substantia nigra. Taking advantage of this unique property of recombinant...

Reversal of motor impairments in parkinsonian rats by continuous intrastriatal delivery of L-dopa using rAAV-mediated gene transfer. (2002)

Kirik, Deniz, Georgievska, Biljana, Burger, Corinna, Winkler, Christian, Muzyczka, Nicholas, Mandel, Ronald J, ...

Intrastriatal delivery of the tyrosine hydroxylase gene by viral vectors is being explored as a tool for local delivery of L-dopa in animals with lesions of the nigrostriatal pathway. The functional...

Cre-mediated recombination in the pituitary gland (2000)

Cushman, Lisa J., Burrows, Heather L., Seasholtz, Audrey F., Lewandoski, Mark, Muzyczka, Nicholas, Camper, Sally A.

Summary: Organ-specific expression of a cre recombinase transgene allows for the analysis of gene function in a particular tissue or cell type. Using a 4.6 kb promoter from the mouse glycoprotein...

A “Humanized ” Green Fluorescent Protein cDNA Adapted for High-Level Expression in Mammalian Cells (1996)

Sergei Zolotukhin, Mark Potter, William W. Hauswirth, John Guy, Nicholas Muzyczka

We constructed gfp h, a synthetic version of the jellyfish Aequorea victoria green fluorescent protein (gfp) cDNA that is adapted for high-level expression in mammalian cells, especially those of...

Efficient photoreceptor-targeted gene expression in vivo by recombinant adeno-associated virus

Flannery, John G., Zolotukhin, Sergei, Vaquero, M. Isabel, LaVail, Matthew M., Muzyczka, Nicholas, Hauswirth, William W.

We describe a general approach for achieving efficient and cell type-specific expression of exogenous genes in photoreceptor cells of the mammalian retina. Recombinant adeno-associated virus (rAAV)...

Sustained secretion of human alpha-1-antitrypsin from murine muscle transduced with adeno-associated virus vectors

Song, Sihong, Morgan, Michael, Ellis, Tamir, Poirier, Amy, Chesnut, Kye, Wang, Jianming, ...

Recombinant adeno-associated virus (AAV) vectors have been used to transduce murine skeletal muscle as a platform for secretion of therapeutic proteins. The utility of this approach for treating...

Mutational Analysis of the Adeno-Associated Virus Type 2 (AAV2) Capsid Gene and Construction of AAV2 Vectors with Altered Tropism

Wu, Pei, Xiao, Wu, Conlon, Thomas, Hughes, Jeffrey, Agbandje-McKenna, Mavis, Ferkol, Thomas, ...

Adeno-associated virus type 2 (AAV2) has proven to be a valuable vector for gene therapy. Characterization of the functional domains of the AAV capsid proteins can facilitate our understanding of...

Biochemical Characterization of Adeno-Associated Virus Rep68 DNA Helicase and ATPase Activities

Zhou, Xiaohuai, Zolotukhin, Irene, Im, Dong-Soo, Muzyczka, Nicholas

The adeno-associated virus (AAV) nonstructural proteins Rep68 and Rep78 are site-specific DNA binding proteins, ATP-dependent site-specific endonucleases, helicases, and ATPases. These biochemical...

Cellular Proteins Required for Adeno-Associated Virus DNA Replication in the Absence of Adenovirus Coinfection

Ni, Tie-Hua, McDonald, William F., Zolotukhin, Irene, Melendy, Thomas, Waga, Shou, Stillman, Bruce, ...

We previously reported the development of an in vitro adeno-associated virus (AAV) DNA replication system. The system required one of the p5 Rep proteins encoded by AAV (either Rep78 or Rep68) and a...

In Vitro Packaging of Adeno-Associated Virus DNA

Zhou, Xiaohuai, Muzyczka, Nicholas

We have developed an in vitro procedure for packaging of recombinant adeno-associated virus (AAV). By using AAV replicative-form DNA as the substrate, it is possible to synthesize an infectious AAV...

Mechanism of Rep-Mediated Adeno-Associated Virus Origin Nicking

Brister, J. Rodney, Muzyczka, Nicholas

The single-stranded adeno-associated virus type 2 (AAV) genome is flanked by terminal repeats (TRs) that fold back on themselves to form hairpinned structures. During AAV DNA replication, the TRs are...

Rep-Mediated Nicking of the Adeno-Associated Virus Origin Requires Two Biochemical Activities, DNA Helicase Activity and Transesterification

Brister, J. Rodney, Muzyczka, Nicholas

The single-stranded adeno-associated virus (AAV) genome is flanked by terminal hairpinned origins of DNA replication (terminal repeats [TRs]) that are nicked at the terminal resolution site (trs) by...

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

Gavin, Denise K., Young, Samuel M., Xiao, Weidong, Temple, Brenda, Abernathy, Corinne R., Pereira, Daniel J., ...

The adeno-associated virus type 2 (AAV) replication (Rep) proteins Rep78 and 68 (Rep78/68) exhibit a number of biochemical activities required for AAV replication, including specific binding to a...

Reversal of motor impairments in parkinsonian rats by continuous intrastriatal delivery of l-dopa using rAAV-mediated gene transfer

Kirik, Deniz, Georgievska, Biljana, Burger, Corinna, Winkler, Christian, Muzyczka, Nicholas, Mandel, Ronald J., ...

Intrastriatal delivery of the tyrosine hydroxylase gene by viral vectors is being explored as a tool for local delivery of l-dopa in animals with lesions of the nigrostriatal pathway. The functional...

Adenovirus-Facilitated Nuclear Translocation of Adeno-Associated Virus Type 2

Xiao, Wu, Warrington, Kenneth H., Hearing, Patrick, Hughes, Jeffrey, Muzyczka, Nicholas

We examined cytoplasmic trafficking and nuclear translocation of adeno-associated virus type 2 (AAV) by using Alexa Fluor 488-conjugated wild-type AAV, A20 monoclonal antibody immunocytochemistry,...

Nigrostriatal α-synucleinopathy induced by viral vector-mediated overexpression of human α-synuclein: A new primate model of Parkinson's disease

Kirik, Deniz, Annett, Lucy E., Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J., Björklund, Anders

We used a high-titer recombinant adeno-associated virus (rAAV) vector to express WT or mutant human α-synuclein in the substantia nigra of adult marmosets. The α-synuclein protein was expressed in...

Identification of Amino Acid Residues in the Capsid Proteins of Adeno-Associated Virus Type 2 That Contribute to Heparan Sulfate Proteoglycan Binding†

Opie, Shaun R., Agbandje-McKenna, Mavis, Zolotukhin, Sergei, Muzyczka, Nicholas

The adeno-associated virus type 2 (AAV2) uses heparan sulfate proteoglycan (HSPG) as its primary cellular receptor. In order to identify amino acids within the capsid of AAV2 that contribute to HSPG...

Circulating Anti-Wild-Type Adeno-Associated Virus Type 2 (AAV2) Antibodies Inhibit Recombinant AAV2 (rAAV2)-Mediated, but Not rAAV5-Mediated, Gene Transfer in the Brain

Peden, Carmen S., Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J.

Epidemiological studies report that 80% of the population maintains antibodies (Ab) to wild-type (wt) adeno-associated virus type 2 (AAV2), with 30% expressing neutralizing Ab (NAb). The blood-brain...

Adeno-Associated Virus Type 2 VP2 Capsid Protein Is Nonessential and Can Tolerate Large Peptide Insertions at Its N Terminus†

Warrington, Kenneth H., Gorbatyuk, Oleg S., Harrison, Jeffrey K., Opie, Shaun R., Zolotukhin, Sergei, Muzyczka, Nicholas

Direct insertion of amino acid sequences into the adeno-associated virus type 2 (AAV) capsid open reading frame (cap ORF) is one strategy currently being developed for retargeting this prototypical...

Structure of Adeno-Associated Virus Type 4

Padron, Eric, Bowman, Valorie, Kaludov, Nikola, Govindasamy, Lakshmanan, Levy, Hazel, Nick, Phillip, ...

Adeno-associated virus (AAV) is a member of the Parvoviridae, belonging to the Dependovirus genus. Currently, several distinct isolates of AAV are in development for use in human gene therapy...

Successful Production of Pseudotyped rAAV Vectors Using a Modified Baculovirus Expression System

Kohlbrenner, Erik, Aslanidi, George, Nash, Kevin, Shklyaev, Stanislav, Campbell-Thompson, Martha, Byrne, Barry J., ...

Scalable production of rAAV vectors remains a major obstacle to the clinical application of this prototypical gene therapy vector. A recently developed baculovirus-based production protocol (M. Urabe...

Studies of the Mechanism of Transactivation of the Adeno-Associated Virus p19 Promoter by Rep Protein

Lackner, Daniel F., Muzyczka, Nicholas

During adeno-associated virus (AAV) type 2 productive infections, the p19 promoter of AAV is activated by the AAV Rep78 and Rep68 proteins. Rep-induced activation of p19 depends on the presence of...

Efficient photoreceptor-targeted gene expression in vivo by recombinant adeno-associated virus

Flannery, John G., Zolotukhin, Sergei, Vaquero, M. Isabel, LaVail, Matthew M., Muzyczka, Nicholas, Hauswirth, William W.

We describe a general approach for achieving efficient and cell type-specific expression of exogenous genes in photoreceptor cells of the mammalian retina. Recombinant adeno-associated virus (rAAV)...

Sustained secretion of human alpha-1-antitrypsin from murine muscle transduced with adeno-associated virus vectors

Song, Sihong, Morgan, Michael, Ellis, Tamir, Poirier, Amy, Chesnut, Kye, Wang, Jianming, ...

Recombinant adeno-associated virus (AAV) vectors have been used to transduce murine skeletal muscle as a platform for secretion of therapeutic proteins. The utility of this approach for treating...

Mutational Analysis of the Adeno-Associated Virus Type 2 (AAV2) Capsid Gene and Construction of AAV2 Vectors with Altered Tropism

Wu, Pei, Xiao, Wu, Conlon, Thomas, Hughes, Jeffrey, Agbandje-McKenna, Mavis, Ferkol, Thomas, ...

Adeno-associated virus type 2 (AAV2) has proven to be a valuable vector for gene therapy. Characterization of the functional domains of the AAV capsid proteins can facilitate our understanding of...

Biochemical Characterization of Adeno-Associated Virus Rep68 DNA Helicase and ATPase Activities

Zhou, Xiaohuai, Zolotukhin, Irene, Im, Dong-Soo, Muzyczka, Nicholas

The adeno-associated virus (AAV) nonstructural proteins Rep68 and Rep78 are site-specific DNA binding proteins, ATP-dependent site-specific endonucleases, helicases, and ATPases. These biochemical...

Cellular Proteins Required for Adeno-Associated Virus DNA Replication in the Absence of Adenovirus Coinfection

Ni, Tie-Hua, McDonald, William F., Zolotukhin, Irene, Melendy, Thomas, Waga, Shou, Stillman, Bruce, ...

We previously reported the development of an in vitro adeno-associated virus (AAV) DNA replication system. The system required one of the p5 Rep proteins encoded by AAV (either Rep78 or Rep68) and a...

In Vitro Packaging of Adeno-Associated Virus DNA

Zhou, Xiaohuai, Muzyczka, Nicholas

We have developed an in vitro procedure for packaging of recombinant adeno-associated virus (AAV). By using AAV replicative-form DNA as the substrate, it is possible to synthesize an infectious AAV...

Mechanism of Rep-Mediated Adeno-Associated Virus Origin Nicking

Brister, J. Rodney, Muzyczka, Nicholas

The single-stranded adeno-associated virus type 2 (AAV) genome is flanked by terminal repeats (TRs) that fold back on themselves to form hairpinned structures. During AAV DNA replication, the TRs are...

Rep-Mediated Nicking of the Adeno-Associated Virus Origin Requires Two Biochemical Activities, DNA Helicase Activity and Transesterification

Brister, J. Rodney, Muzyczka, Nicholas

The single-stranded adeno-associated virus (AAV) genome is flanked by terminal hairpinned origins of DNA replication (terminal repeats [TRs]) that are nicked at the terminal resolution site (trs) by...

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

Gavin, Denise K., Young, Samuel M., Xiao, Weidong, Temple, Brenda, Abernathy, Corinne R., Pereira, Daniel J., ...

The adeno-associated virus type 2 (AAV) replication (Rep) proteins Rep78 and 68 (Rep78/68) exhibit a number of biochemical activities required for AAV replication, including specific binding to a...

Reversal of motor impairments in parkinsonian rats by continuous intrastriatal delivery of l-dopa using rAAV-mediated gene transfer

Kirik, Deniz, Georgievska, Biljana, Burger, Corinna, Winkler, Christian, Muzyczka, Nicholas, Mandel, Ronald J., ...

Intrastriatal delivery of the tyrosine hydroxylase gene by viral vectors is being explored as a tool for local delivery of l-dopa in animals with lesions of the nigrostriatal pathway. The functional...

Adenovirus-Facilitated Nuclear Translocation of Adeno-Associated Virus Type 2

Xiao, Wu, Warrington, Kenneth H., Hearing, Patrick, Hughes, Jeffrey, Muzyczka, Nicholas

We examined cytoplasmic trafficking and nuclear translocation of adeno-associated virus type 2 (AAV) by using Alexa Fluor 488-conjugated wild-type AAV, A20 monoclonal antibody immunocytochemistry,...

Nigrostriatal α-synucleinopathy induced by viral vector-mediated overexpression of human α-synuclein: A new primate model of Parkinson's disease

Kirik, Deniz, Annett, Lucy E., Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J., Björklund, Anders

We used a high-titer recombinant adeno-associated virus (rAAV) vector to express WT or mutant human α-synuclein in the substantia nigra of adult marmosets. The α-synuclein protein was expressed in...

Studies of the Mechanism of Transactivation of the Adeno-Associated Virus p19 Promoter by Rep Protein

Lackner, Daniel F., Muzyczka, Nicholas

During adeno-associated virus (AAV) type 2 productive infections, the p19 promoter of AAV is activated by the AAV Rep78 and Rep68 proteins. Rep-induced activation of p19 depends on the presence of...

Identification of Amino Acid Residues in the Capsid Proteins of Adeno-Associated Virus Type 2 That Contribute to Heparan Sulfate Proteoglycan Binding†

Opie, Shaun R., Agbandje-McKenna, Mavis, Zolotukhin, Sergei, Muzyczka, Nicholas

The adeno-associated virus type 2 (AAV2) uses heparan sulfate proteoglycan (HSPG) as its primary cellular receptor. In order to identify amino acids within the capsid of AAV2 that contribute to HSPG...

Circulating Anti-Wild-Type Adeno-Associated Virus Type 2 (AAV2) Antibodies Inhibit Recombinant AAV2 (rAAV2)-Mediated, but Not rAAV5-Mediated, Gene Transfer in the Brain

Peden, Carmen S., Burger, Corinna, Muzyczka, Nicholas, Mandel, Ronald J.

Epidemiological studies report that 80% of the population maintains antibodies (Ab) to wild-type (wt) adeno-associated virus type 2 (AAV2), with 30% expressing neutralizing Ab (NAb). The blood-brain...

Adeno-Associated Virus Type 2 VP2 Capsid Protein Is Nonessential and Can Tolerate Large Peptide Insertions at Its N Terminus†

Warrington, Kenneth H., Gorbatyuk, Oleg S., Harrison, Jeffrey K., Opie, Shaun R., Zolotukhin, Sergei, Muzyczka, Nicholas

Direct insertion of amino acid sequences into the adeno-associated virus type 2 (AAV) capsid open reading frame (cap ORF) is one strategy currently being developed for retargeting this prototypical...

Structure of Adeno-Associated Virus Type 4

Padron, Eric, Bowman, Valorie, Kaludov, Nikola, Govindasamy, Lakshmanan, Levy, Hazel, Nick, Phillip, ...

Adeno-associated virus (AAV) is a member of the Parvoviridae, belonging to the Dependovirus genus. Currently, several distinct isolates of AAV are in development for use in human gene therapy...

Structurally Mapping the Diverse Phenotype of Adeno-Associated Virus Serotype 4▿

Govindasamy, Lakshmanan, Padron, Eric, McKenna, Robert, Muzyczka, Nicholas, Kaludov, Nikola, Chiorini, John A., ...

The adeno-associated viruses (AAVs) can package and deliver foreign DNA into cells for corrective gene delivery applications. The AAV serotypes have distinct cell binding, transduction, and antigenic...

Production, purification, crystallization and preliminary X-ray analysis of adeno-associated virus serotype 8

Lane, Michael Douglas, Nam, Hyun-Joo, Padron, Eric, Gurda-Whitaker, Brittney, Kohlbrenner, Eric, Aslanidi, George, ...

The production, purification, crystallization and preliminary X-ray crystallographic analysis of adeno-associated virus serotype 8 is reported.

Purification of Host Cell Enzymes Involved in Adeno-Associated Virus DNA Replication▿

Nash, Kevin, Chen, Weijun, McDonald, William F., Zhou, Xiaohuai, Muzyczka, Nicholas

Adeno-associated virus (AAV) replicates its DNA by a modified rolling-circle mechanism that exclusively uses leading strand displacement synthesis. To identify the enzymes directly involved in AAV...

Production, purification, crystallization and preliminary X-ray structural studies of adeno-associated virus serotype 5

DiMattia, Michael, Govindasamy, Lakshmanan, Levy, Hazel C., Gurda-Whitaker, Brittney, Kalina, Amy, Kohlbrenner, Erik, ...

The production, purification, crystallization and preliminary crystallographic analysis of empty adeno-associated virus serotype 5 capsids are reported.

Structure of Adeno-Associated Virus Serotype 8, a Gene Therapy Vector▿

Nam, Hyun-Joo, Lane, Michael Douglas, Padron, Eric, Gurda, Brittney, McKenna, Robert, Kohlbrenner, Erik, ...

Adeno-associated viruses (AAVs) are being developed as gene therapy vectors, and their efficacy could be improved by a detailed understanding of their viral capsid structures. AAV serotype 8 (AAV8)...

Complete In Vitro Reconstitution of Adeno-Associated Virus DNA Replication Requires the Minichromosome Maintenance Complex Proteins▿

Nash, Kevin, Chen, Weijun, Muzyczka, Nicholas

Adeno-associated virus (AAV) replicates its DNA exclusively by a leading-strand DNA replication mechanism and requires coinfection with a helper virus, such as adenovirus, to achieve a productive...

The phosphorylation state of Ser-129 in human α-synuclein determines neurodegeneration in a rat model of Parkinson disease

Gorbatyuk, Oleg S., Li, Shoudong, Sullivan, Layla F., Chen, Weijun, Kondrikova, Galina, Manfredsson, Fredric P., ...

Studies have shown that α-synuclein (α-syn) deposited in Lewy bodies in brain tissue from patients with Parkinson disease (PD) is extensively phosphorylated at Ser-129. We used recombinant...

Production, purification and preliminary X-ray crystallographic studies of adeno-associated virus serotype 1

Miller, Edward B., Gurda-Whitaker, Brittney, Govindasamy, Lakshmanan, McKenna, Robert, Zolotukhin, Sergei, Muzyczka, Nicholas, ...

Crystals of baculovirus-expressed adeno-associated virus serotype 1 (AAV1) capsids have been grown in the rhombohedral space group R32 (unit-cell parameters a = 254.7 Å, α = 62.3°) and shown to...

Next generation of adeno-associated virus 2 vectors: Point mutations in tyrosines lead to high-efficiency transduction at lower doses

Zhong, Li, Li, Baozheng, Mah, Cathryn S., Govindasamy, Lakshmanan, Agbandje-McKenna, Mavis, Cooper, Mario, ...

Recombinant adeno-associated virus 2 (AAV2) vectors are in use in several Phase I/II clinical trials, but relatively large vector doses are needed to achieve therapeutic benefits. Large vector doses...

Identification of Cellular Proteins That Interact with the Adeno-Associated Virus Rep Protein▿

Nash, Kevin, Chen, Weijun, Salganik, Max, Muzyczka, Nicholas

Adeno-associated virus (AAV) codes for four related nonstructural Rep proteins. AAV both replicates and assembles in the nucleus and requires coinfection with a helper virus, either adenovirus (Ad)...