Gene delivery system for retina research based on recombinant adenoassociated viral vector
会议名称：《中国神经科学学会第四次会员代表大会暨第七届全国学术会议》
会议日期：2007年
学科分类：0710[理学-生物学] 07[理学] 071003[理学-生理学]
关 键 词：gene delivery system retina adeno-associated virus(AAV) bicitronic vector
摘 要：正Purpose To establish an efficient and stable gene delivery system based on recombinant adeno-associated viral vectors for retina research,such as dendrite development patterns of mouse retinal cells,especially retinal ganglion cells, and how the cell fate of various retinal cells is *** A bicistronic plasmid vector,pSIEWB,was acquired by molecular manufacture with elements derived from pSEWB which contains AAV genome and provides main structure of the vector and pCAGGS which provides the fragment containing IRES and EGFP *** of recombinant AAV vector was performed by a dual-vector,calcium phosphate-mediated transfection approach with pSIEWB and pD1rs which provides the AAV coat protein genes and helper virus genes necessary for packaging of recombinant AAV vector. Results The bicistronic plasmid vector,pSIEWB,has been acquired via replacing EGFP gene of pSEWB by fragment containing IRES-EGFP sequence derived from pCAGGS and it possesses the genome of recombinant AAV including two inverted terminal repeat elements（ITRs） which are necessary for viral packaging,human synapsin 1 promoter（hSyn 1） which can drive gene expression in neuron specially,IRES which can make functional gene and reporter gene express individually,EGFP which is a popular reporter gene to localize gene expression of vector,and woodchuck hepatitis post-transcriptional regulatory element（WPRE） which can optimize gene expression in *** the same time,an dual- vector, calcium phosphate-mediated transfection approach for viral packaging has been performed efficiently and stably in our lab,which needs two plasmids,pSIEWB and pD1rs which works as a helper plasmid for viral packaging because it contains AAV coat protein genes and adenovirus genes necessary for *** This study has provided a primary form of a gene delivery system for retina *** study on the process of dendrite development of retinal ganglion cells and cell fate determinatio