title: Distinct Expression Patterns of AAV8 Vectors with Broadly Active Promoters from Subretinal Injections of Neonatal Mouse Eyes at Two Different Ages.
abstract: The retinal expression patterns were analyzed following the injection of serotype 8 adeno-associated virus (AAV8) vectors that utilize two broadly active and commonly used sets of transcription regulatory sequences. These include the human cytomegalovirus (CMV) immediate early (IE) enhancer/promoter and the hybrid CAG element (also known as CAGGS or CBA) composed of a partial human CMV IE enhancer and the chicken β-actin promoter and intron. Subretinal delivery to postnatal day 0 (P0) or 6 (P6) mouse eyes resulted in efficient labeling of retinal cells, but with very distinct patterns. With P0 delivery, AAV8-CMV-GFP selectively labelled photoreceptors, while AAV8-CAG-GFP efficiently labeled both outer and inner retinal neurons, including photoreceptors, horizontal cells, amacrine cells and retinal ganglion cells. With P6 delivery, both vectors led to efficient labeling of photoreceptors and Müller glia cells, but not of inner retinal neurons. Our results suggest that the cell types that express the genes encoded by subretinally delivered AAV8 vectors are determined by both the timing of the injection and the regulatory sequences.
authors.affiliation: Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Harvard Medical School, 77 Avenue Louis Pasteur, NRB 360, 02115, Boston, MA, USA. wjxiong@genetics.med.harvard.edu.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Xiong
authors.affiliation: Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Harvard Medical School, 77 Avenue Louis Pasteur, NRB 360, 02115, Boston, MA, USA. cepko@genetics.med.harvard.edu.
authors.forename: Constance
authors.initials: C
authors.lastname: Cepko
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000229:Dependovirus / Q000235:genetics*; D020869:Gene Expression Profiling; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D007267:Injections; D051379:Mice; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012160:Retina / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic
chemical_list: D049452:Green Fluorescent Proteins
keywords: AAV8; Cellular tropism; Chicken β-actin promoter/enhancer/intron; Human cytomegalovirus (CMV) immediate early (IE) enhancer/promoter; Neonatal mouse eye; Subretinal injection; Transgene expression