Presence of a retroviral encapsidation sequence in nonretroviral RNA increases the efficiency of formation of cDNA genes.
We showed previously that retrovirus vector particles can encapsidate RNAs without retroviral cis-acting sequences, that such RNAs are reverse transcribed in infected target cells, and that the cDNA copies are inserted into the host genome resulting in cDNA genes (R. Dornburg and H. M. Temin, Mol. Cell. Biol. 8:2328-2334, 1988). To provide further evidence that this retrovirus-mediated gene transfer occurred through an RNA intermediate, we constructed retroviral vectors containing an intron from a cellular gene. This intron was lost in a cDNA gene formed after infection with retroviral particles, establishing that an RNA intermediate had existed. Retroviral vectors with additional encapsidation sequences were constructed. The presence of a murine leukemia virus encapsidation sequence in an mRNA transcribed from the hygromycin B phosphotransferase gene increased the efficiency of encapsidation into spleen necrosis virus vector particles and the formation of cDNA genes by approximately 2 orders of magnitude.
Dornburg
R
R
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Temin
H M
HM
D000818:Animals; D002213:Capsid / Q000235:genetics*; D002460:Cell Line; D005796:Genes; D005809:Genes, Regulator; D005814:Genes, Viral; D005822:Genetic Vectors; D007438:Introns; D009052:Leukemia Virus, Murine / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011533:Proviruses / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics; D006377:T-Lymphocytes, Helper-Inducer; D013937:Thymidine Kinase / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics
D015678:Viral Structural Proteins; D013937:Thymidine Kinase