Analysis of gene function in the retina.
The Retina is a good model system for studies of neural development and disease because of its simplicity and accessibility. To analyze gene function rapidly and conveniently, we developed an electroporation technique in mice and rats for use in vivo and in vitro. The efficiency of electroporation into the neonatal retina is quite good, and transgene expression persists for more than a month. With this technique, various types of DNA constructs, including RNA interference (RNAi) vectors, are readily introduced into the retina without DNA size limitation. In addition, more than two different DNA constructs can be introduced into the retina at once, with very high cotransfection efficiency. In vivo and in vitro electroporation will provide a powerful method to analyze the molecular mechanisms of retinal development and disease.
Matsuda
Takahiko
T
Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA.
Cepko
Constance L
CL
D000818:Animals; D000831:Animals, Newborn; D018274:Electroporation / Q000379:methods*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D066298:In Vitro Techniques; D051379:Mice; D008959:Models, Neurological; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics; D012160:Retina / Q000254:growth & development / Q000378:metabolism*
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins