General Utility of the Chicken βB1-Crystallin Promoter to Drive Protein Expression in Lens Fiber Cells of Transgenic Mice
pmid: 12212842
General Utility of the Chicken βB1-Crystallin Promoter to Drive Protein Expression in Lens Fiber Cells of Transgenic Mice
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 alphaA-crystallin promoter. However, while lens-specific, transgenic lines made with the alphaA-crystallin promoter express the transgene at levels 100-300-fold lower than endogenous alphaA-crystallin. Here we propose an alternative, the chicken betaB1-crystallin promoter (-432/+30). Transgenic mice made with this promoter have successfully expressed CAT, d/n m-calpain, Weel, and betaB2-crystallin mRNA at levels comparable to the endogenous betaB1-crystallin gene and no eye abnormalities such as cataracts, have resulted. All of the transgenic lines made with the chicken betaB1-crystallin promoter have expressed the transgene in the lens fiber cells, and the best lines express at levels close to endogenous betaB1-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 betaB1-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.
- National Institutes of Health United States
- Oregon Health & Science University United States
- National Eye Institute United States
- University of Delaware United States
- National Institute of Health Pakistan
Base Sequence, Molecular Sequence Data, Mice, Transgenic, Crystallins, Polymerase Chain Reaction, Mice, Sequence Homology, Nucleic Acid, Consensus Sequence, Lens, Crystalline, beta-Crystallin B Chain, Animals, Humans, Electrophoresis, Gel, Two-Dimensional, Promoter Regions, Genetic, Chickens, Sequence Alignment, DNA Primers
Base Sequence, Molecular Sequence Data, Mice, Transgenic, Crystallins, Polymerase Chain Reaction, Mice, Sequence Homology, Nucleic Acid, Consensus Sequence, Lens, Crystalline, beta-Crystallin B Chain, Animals, Humans, Electrophoresis, Gel, Two-Dimensional, Promoter Regions, Genetic, Chickens, Sequence Alignment, DNA Primers
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