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Robert M. Kotin, Ph.D.

Laboratory of Molecular Virology and Gene Therapy

Robert M. Kotin, Ph.D.
Senior Investigator
Laboratory of Molecular Virology and Gene Therapy

Building 10 Room 7D05
10 Center Dr
Bethesda, MD 20892
P: +1 301 496 1594
F: +1 301 496 9985


Robert Kotin graduated from the University of California, Santa Cruz with a B.A. in biology in 1978 and from Rutgers University and the University of Medicine and Dentistry of New Jersey with a Ph.D. in microbiology in 1986. He was a Norman and Rosita Winston Foundation Research Fellow from 1986 to 1988 and a research associate at Cornell University Medical College from 1989 to 1990. Dr. Kotin joined the NHLBI in 1994 as a tenure-track Investigator and was promoted to Senior Investigator in 1999. Dr. Kotin has authored or coauthored more than 75 papers and is an inventor on ten U.S. issued patents. He is an associate editor for the Journal of Gene Medicine, and on the editorial boards of Gene Therapy and Human Gene Therapy.

Research Interests

The characteristics that make viruses such a threat to human health also make them a vital tool in the development of gene therapies. Dr. Kotin discovered that in cells latently infected with adeno-associated virus (AAV) the viral DNA was frequently integrated at a common site on human chromosome 19, which he designated as locus AAVS1. This phenomenon appears unique to AAV among animal viruses and may have utility for gene transfer. The trick is to understand mechanistically how AAV acts in order to exploit this valuable machinery. Dr. Kotin’s laboratory focuses on understanding the molecular biology of AAV, a widespread human parvovirus, to develop AAV as a gene transfer vector for the treatment of human disease.

Recombinant AAV (rAAV) vectors have been used in tissue culture and small animal experiments with great success; however, their clinical impact has been limited. One practical difficulty has been the production of sufficient quantities of clinical and research grade rAAV to obtain meaningful results in clinically relevant models of human diseases. In recent years, Dr. Kotin has been heavily involved in developing the methodologies to meet this need. Dr. Kotin and members of his laboratory discovered that they could produce this mammalian virus in an insect cell system, Spodoptera frugiperda (Sf9 cells), with a much higher efficiency than possible in the widely used mammalian HEK 293 cells. The Sf9 cells grow in suspension cultures allowing the process to be scaled-up volumetrically. The highly scalable production system that resulted from this research has been licensed by several companies pursuing gene therapies, leading to the first AAV gene therapy recommended for marketing approval by the European Medicines Agency.

Dr. Kotin is applying this system to generate a gene therapy approach for Duchenne muscular dystrophy (DMD), the most common form of hereditary myopathy that affects every striated muscle in the body. In addition to increasing the efficiency and capacity of rAAV production, Dr. Kotin is also pursuing strategies to study different AAV serotypes, with different biological properties, that cannot be purified from cell lysates using commercially available chromatography media. His laboratory recently developed a unique affinity column and derived a recombinant single chain antibody that is highly specific for AAV9 and is working through a cooperative research and development agreement to make the resource commercially available. And very recently, he has found a novel way to produce closed-end, linear duplex (CELiD) DNA in eukaryotic cells as an alternative to plasmid DNA obtained from bacteria as a way to introduce genetic information that does not integrate into the host cell.

His enthusiasm for solving biotechnology problems to speed the development of AAV-based gene therapies notwithstanding, Dr. Kotin continues to take a deep interest in understanding the basic biological mechanisms of viral infection. AAV is a dependovirus, that is, it requires a nuclear DNA virus that co-infects its host. In developing the Sf9 viral expression system, his team discovered that a baculovirus provides the necessary and sufficient genes to allow co-infection by AAV. He is currently studying the baculovirus genes that are critical for successful co-infection. Once uncovered, these critical Baculoviridae genes can be compared to previously characterized co-infection genes from two other DNA virus types, the Adenoviridae and Herpesviridae. This triangulation can provide the means to better understand the nature and evolution of the helper virus.

Selected Publications

MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy.
Barbash IM, Cecchini S, Faranesh AZ, Virag T, Li L, Yang Y, Hoyt RF, Kornegay JN, Bogan JR, Garcia L, Lederman RJ, Kotin RM.
Gene Ther. 2012 May 3.
[Text Abstract on PubMed]

Reproducible high yields of recombinant adeno-associated virus produced using invertebrate cells in 0.02- to 200-liter cultures.
Cecchini S, Virag T, Kotin RM.
Hum. Gene Ther. 2011 Aug;22(8):1021-30.
[Text Abstract on PubMed]

A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells.
Smith RH, Levy JR, Kotin RM.
Mol. Ther. 2009 Nov;17(11):1888-96.
[Text Abstract on PubMed]

Insect cells as a factory to produce adeno-associated virus type 2 vectors.
Urabe M, Ding C, Kotin RM.
Hum. Gene Ther. 2002 Nov 1;13(16):1935-43.
[Text Abstract on PubMed]

Site-specific integration by adeno-associated virus.
Kotin RM, Siniscalco M, Samulski RJ, Zhu XD, Hunter L, Laughlin CA, McLaughlin S, Muzyczka N, Rocchi M, Berns KI.
Proc. Natl. Acad. Sci. U.S.A. 1990 Mar;87(6):2211-5.
[Text Abstract on PubMed]

Robert M. Kotin's Full List of Publications

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Last Updated: March 31, 2014

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