Sequences in pol are required for transfer of human foamy virus-based vectors.
A series of vectors with heterologous genes was constructed from HSRV1, an infectious clone of human foamy virus (HFV), and transfected into baby hamster kidney cells to generate stably transfected vector cell lines. Two cis-acting sequences were required to achieve efficient rescue by helper virus. The first element was located at the 5' end upstream of position 1274 of the proviral DNA. Interestingly, a mutation in the leader sequence which decreased the ability to dimerize in vitro inhibited transfer by helper HFV. A second element that was important for vector transfer was located in the pol gene between positions 5638 and 6317. Constructs lacking this element were only poorly transferred by helper HFV, even though their RNA was produced in the vector cell lines. This finding rules out the possibility that the observed lack of transfer was due to RNA instability. A minimal vector containing only these two elements could be successfully delivered by helper HFV, confirming that all essential cis-acting sequences were present. The presence of a sequence described as a second polypurine tract in HFV was not necessary for transfer. Our data identified the minimal sequence requirements for HFV vector transfer for the development of useful vector systems.
Erlwein
O
O
Department of Genito-Urinary Medicine and Communicable Diseases, Jefferiss Research Trust Laboratories, Imperial College School of Medicine at St. Mary's, London W2 1NY, United Kingdom.
Bieniasz
P D
PD
McClure
M O
MO
D000818:Animals; D001483:Base Sequence; D006224:Cricetinae; D018014:Gene Transfer Techniques; D015751:Genes, pol; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D012367:RNA, Viral / Q000096:biosynthesis; D016092:Spumavirus / Q000235:genetics*
D012367:RNA, Viral