Stable expression of three genes from a tricistronic retroviral vector containing a picornavirus and 9-nt cellular internal ribosome entry site elements.
Retroviral vectors are used widely to deliver heterologous genes into cells. In order to express three genes from a single RNA molecule, a retroviral vector that contains two divergent internal ribosome entry site (IRES) sequences has been constructed successfully. To eliminate the high frequency of recombination within a mulicistronic retrovirus vector, a 9-nt segment of a cellular mRNA IRES and a picornaviral IRES were used, since these two IRES sequences have minimal sequence homology. After a single round of replication, most cells infected with this vector stably expressed the three genes while approximately 40% of cells infected with another tricistronic retroviral vector that contains two copies of an identical IRES sequence lost expression of the gene located between these two sequences.
Li
Ting
T
Department of Microbiology and Immunology and Markey Cancer Center, University of Kentucky, 206 Combs Research Building, 800 Rose Street, Lexington, KY 40536-0096, USA.
Zhang
Jiayou
J
D001483:Base Sequence; D002460:Cell Line; D004686:Encephalomyocarditis virus / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D014773:Virology / Q000379:methods
D012333:RNA, Messenger; D012367:RNA, Viral