General utility of the chicken betaB1-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.
Taube
Jennifer R
JR
Department of Biological Sciences, University of Delaware, Newark 19716, USA.
Gao
Chun Y
CY
Ueda
Yoji
Y
Zelenka
Peggy S
PS
David
Larry L
LL
Duncan
Melinda K
MK
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D016384:Consensus Sequence; D003459:Crystallins / Q000235:genetics*; D017931:DNA Primers; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007908:Lens, Crystalline / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D038206:beta-Crystallin B Chain
C105601:CRYBB1 protein, human; C491363:Crybb1 protein, mouse; D003459:Crystallins; D017931:DNA Primers; D038206:beta-Crystallin B Chain