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General Utility of the Chicken βB1-Crystallin Promoter to Drive Protein Expression in Lens Fiber Cells of Transgenic Mice

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Abstract

Transgenic mouse technology has been very valuable for the study of lens fiber cells since they can not be propagated in cell culture. The targeting of transgenes to the lens has traditionally been done with the αA-crystallin promoter. However, while lens-specific, transgenic lines made with the αA-crystallin promoter express the transgene at levels 100–300-fold lower than endogenous αA-crystallin. Here we propose an alternative, the chicken βB1-crystallin promoter (−432/+30). Transgenic mice made with this promoter have successfully expressed CAT, d/n m-calpain, Wee1, and βB2-crystallin mRNA at levels comparable to the endogenous βB1-crystallin gene and no eye abnormalities such as cataracts, have resulted. All of the transgenic lines made with the chicken βB1-crystallin promoter have expressed the transgene in the lens fiber cells, and the best lines express at levels close to endogenous βB1-crystallin. While RNA expression is very high, only moderate protein expression has been achieved, implying that the high protein expression of the crystallins is partially controlled at the level of translation. Thus, the chicken βB1-crystallin promoter directs high level RNA expression to lens fiber cells, which may be especially useful for the expression of ribozyme and anti-sense RNAs in addition to ectopic proteins.

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Author notes

  1. Yoji Ueda

    Present address: TRA Urology, Research Center Kyoto, Bayer Yakuhin, Ltd., 6-5-1-3, Kunimidai, Kizu-cho, Soraku-gun, Kyoto, 619-0216, Japan

Authors and Affiliations

  1. Department of Biological Sciences, University of Delaware, 314 Wolf Hall, Newark, DE, 19716, USA

    Jennifer R. Taube & Melinda K. Duncan

  2. Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, MD, 20892-2730, USA

    Chun Y. Gao & Peggy S. Zelenka

  3. Departments of Oral Molecular Biology and Ophthalmology, Schools of Dentistry and Medicine, Oregon Health Sciences University, Portland, OR, 97201, USA

    Yoji Ueda & Larry L. David

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  1. Jennifer R. Taube
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Correspondence to Melinda K. Duncan.

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Taube, J.R., Gao, C.Y., Ueda, Y. et al. General Utility of the Chicken βB1-Crystallin Promoter to Drive Protein Expression in Lens Fiber Cells of Transgenic Mice. Transgenic Res 11, 397–410 (2002). https://doi.org/10.1023/A:1016364001095

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