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S.L. Boye, J.J. Peterson, H. Petrs–Silva, S.–H. Min, J. Pang, S. Haire, Q. Li, V.A. Chiodo, M. Ding, W.W. Hauswirth; Transduction and Tropism of an Abbreviated Form of CMV–Chicken ß–Actin Promoter (CBA) With AAV in Mouse Retina . Invest. Ophthalmol. Vis. Sci. 2006;47(13):852.
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The chimeric CMV–chicken ß–actin promoter (CBA) has been utilized extensively as a promoter that supports expression in a wide variety of cells when in recombinant adeno–associated virus (AAV) vectors delivered to retina. Given the limited packaging capacity of AAV, large size of standard CBA and the recent development of self–complementary AAV vectors (further limited in size) an abbreviated form of the CBA would have wide utility.
In these studies we directly compare full length CBA (1720 bases) to a smaller version, smCBA (953 bases) which has the hybrid chicken ß–actin/rabbit ß–globin intron greatly shortened. Experiments used AAV type 2 and 5 and green fluorescent protein as the reporter gene. Both subretinal and intravitreal injections were done in mice. We also used smCBA to express transgenes in two disease models and evaluated rescue. Finally, we combined smCBA with self–complementary AAV (scAAV) vectors, taking advantage of the broad tropism of CBA coupled with the increased transduction speed of self–complementary AAV.
Experiments comparing the trophism of smCBA to full CBA indicate identical patterns of expression. Both subretinal and intravitreal injections of vector resulted in the same general level of expression and cell types targeted. In two disease models, the RD10 mouse and the GC1 knock–out mouse, rescue was demonstrated using AAV5–smCBA driving the respective therapeutic transgene. Our lab is currently evaluating several self–complementary AAV vectors using smCBA. These include therapeutic approaches to diabetic retinopathy.
An abbreviated form of the CBA promoter works well in vivo when packaged into AAV vectors. Its use in AAV vectors would allow larger cDNAs to be tested for therapeutic efficacy than are currently feasible.
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