Transgenics and Genome Manipulation Section

Building 7, Room 104 7 Memorial Drive, MSC 0704 Bethesda, Maryland 20892-0704 Phone: (301) 496-8841 Fax: (301) 402-3603

• Activities/Accomplishments
• Staff
• Recent Publications

Activities/Accomplishments

This section is composed of two sub-sections, 1. a research subunit and 2. the NEI Central Transgenic Facility. The mission of the NEI Central Transgenic Facility is to provide transgenic technology and expertise to researchers in the NEI intramural research program. This includes collaborating on design of transgene constructs, generating and maintaining transgenic mouse lines for NEI researchers, analyzing their DNA for presence of the desired transgene, cryopreserving important lines, maintaining accurate, up-to-date computer records on all transgenic mice, and collaborating on creation of gene knockout mice.

The research subunit is investigating the in vivo functions of the lens α-crystallin proteins (αA and αB). These proteins, abundant in vertebrate lenses, were originally thought to be solely structural proteins, but in recent years have been shown to posses a myriad of activities in vitro including molecular chaperone activity, autokinase activity, DNA binding activity and binding to and regulating the polymerization state of several cytoskeletal proteins. Mice lacking the α-crystallins have been generated and are being analyzed. Mice lacking αB-crystallin appear to be completely normal. Mice lacking αA however develop cataracts beginning early in life and progressing in severity with age. In the absence of αA, the αB forms large insoluble masses, 1 to 3 um in diameter, in the cytoplasm in lens fiber cells. Lenses of mice lacking αB-crystallin appear to be completely normal. However, these mice exhibit dystrophy of select skeletal muscles, particularly those of the head (especially tongue) and those surrounding the spine. Mice lacking both α-crystallins exhibit a severe cataract very early in life and also develop the muscular dystrophy observed in αB-crystallin knockout mice.

Staff

Name	Title	E-mail	Phone
Eric Wawrousek, Ph.D.	Section Head	ericw@helix.nih.gov	(301) 496-8841
R. Steven Lee	Biologist	slee@helix.nih.gov	(301) 402-2728
Carl Haugen	Biologist	Haugenc@intra.nei.nih.gov	(301) 402-2728

Recent Publications

Nakayama M, Stauffer J, Cheng J, Banerjee-Basu S, Wawrousek E and Buonanno A (1996) Common Core Sequences are Found in Skeletal Muscle Slow- and Fast-Fiber-Type-Specific Regulatory Elements, Mol. Cell. Biol., 16(5), 2408.

Brady JP, Garland D, Duglas-Tabor Y, Robison WG Jr, Groome A and Wawrousek EF (1997) Targeted Disruption of the Mouse αA-Crystallin Gene Induces Cataract and Cytoplasmic Inclusion Bodies Containing the Small Heat Shock Protein αB-Crystallin, Proc. Natl. Acad. Sci. USA. 94, 884.

Sunayashiki-Kusuzaki K, Kikuchi T, Wawrousek EF and Shinohara T (1997) Arrestin and Phosducin are Expressed in a Small Number of Brain Cells, Molecular Brain Research, 52, 112.

Lai JC, Fukushima A, Wawrousek EF, Lobanoff MC, Charukamnoetkanok P, Smith-Gill SJ, Vistica BP, Lee RS, Egwuagu CE, Whitcup SM, Gery I (1998) Immunotolerance Again st a Foreign Antigen Transgenically Expressed in the Lens, Invest. Ophthalmol. Vis. Sci., 39(11), 2049-2057.

Andley UP, Song Z, Wawrousek EF, and Bassnett S (1998) The Molecular Chaperone αA-Crystallin Enhances Lens Epithelial Cell Growth and Resistance to UVA Stress, J. Biol. Chem., 273(47), 31252-31261.

Lai JC, Lobanoff MC, Fukushima A, Wawrousek EF, Chan CC, Whitcup SM, Gery I (1999) Uveitis Induced by Lymphocytes Sensitized Against a Transgenically Expressed Lens Protein, Invest. Ophthalmol. Vis. Sci., 40(11), 2735-2739.

Xu H, Wawrousek EF, Redmond TM, Nickerson JM, Wiggert B, Chan CC, Caspi RR (2000) Transgenic Expression of an Immunologically Privileged Retinal Antigen Extraocularly Enhances Self Tolerance and Abrogates Susceptibility to Autoimmune Uveitis, Eur. J. Immunol., 30(1), 272-278.

Kimura A, Singh D, Wawrousek EF, Kikuchi M, Nakamura M, Shinohara T (2000) Both PCE-1/RX and OTX/CRX Interactions are Necessary for Photoreceptor-specific Gene Expression, J. Biol. Chem. 275(2), 1152-1160.