Herweijer H, Latendresse J S, Williams P, Zhang G, Danko I, Schlesinger S, Wolff J A
Department of Pediatrics and Medical Genetics, University of Wisconsin, Madison 53705, USA.
Hum Gene Ther. 1995 Sep;6(9):1161-7. doi: 10.1089/hum.1995.6.9-1161.
Sindbis virus was used as a self-amplifying eukaryotic expression vector. A recombinant cDNA genome of this (+)-strand RNA virus was placed under the transcriptional control of a Rous sarcoma virus LTR (RSV) promoter. Transfection of this plasmid construct into mammalian cell lines (3T3, HepG2, and 293 cells) resulted in expression of the luciferase reporter gene. High-expression levels were also measured after transfection into primary rat myoblasts. In differentiated myotubes, expression levels generated by the Sindbis virus vector were up to 200 times higher than those obtained with a conventional RSV expression vector. In vivo expression was detected after injection of plasmid DNA into mouse quadriceps. In vivo expression was transient and undetectable by day 16. This self-amplifying expression vector can be used for generating high-level expression of transgenes in vitro and in vivo. Its transient nature in vivo could allow for safe, short-term delivery of gene products in gene therapy protocols. It should facilitate the study of Sindbis and other RNA viruses.
辛德毕斯病毒被用作一种自我扩增的真核表达载体。这种正链RNA病毒的重组cDNA基因组置于劳氏肉瘤病毒长末端重复序列（RSV）启动子的转录控制之下。将该质粒构建体转染到哺乳动物细胞系（3T3、HepG2和293细胞）中导致荧光素酶报告基因的表达。转染到原代大鼠成肌细胞后也检测到高表达水平。在分化的肌管中，辛德毕斯病毒载体产生的表达水平比用传统RSV表达载体获得的表达水平高200倍。将质粒DNA注射到小鼠股四头肌后检测到体内表达。体内表达是短暂的，到第16天时无法检测到。这种自我扩增的表达载体可用于在体外和体内产生转基因的高水平表达。其在体内的短暂性质可允许在基因治疗方案中安全、短期地递送基因产物。它应有助于对辛德毕斯病毒和其他RNA病毒的研究。
