Ultrasound-guided in utero injections allow studies of the development and function of the eye.
Ultrasound-guided in utero injections into the brain of murine embryos has been shown to facilitate gene delivery. We investigated whether these methods would allow gene transfer into ocular structures. Gene transfer using retroviral vectors or electroporation was found to be quite effective. We determined the window of time, as well as compared several strains of mice, that yield a high degree of survival and successful gene transfer. Several retroviral constructs were tested for expression and coexpresssion of two genes in retinal cell types. In addition, a retroviral vector was engineered to give cone photoreceptor-enriched expression, and a retroviral vector was demonstrated to provide RNAi-mediated loss-of-function. These methods enable access to early ocular structures and provide a more rapid method of assessment of gene and promoter function than possible using genetically engineered mice.
Punzo
Claudio
C
Harvard Medical School, Department of Genetics, Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA.
Cepko
Constance L
CL
D000818:Animals; D018274:Electroporation; D004622:Embryo, Mammalian / Q000033:anatomy & histology / Q000000981:diagnostic imaging / Q000502:physiology; D005123:Eye / Q000000981:diagnostic imaging / Q000196:embryology / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D007267:Injections; D051379:Mice; D008815:Mice, Inbred Strains; D014463:Ultrasonography