Generation of a helper cell line for packaging avian leukosis virus-based vectors.
We constructed an avian leukosis virus-based packaging cell line, pHF-g, containing Rous-associated virus DNA with several alterations to abolish RNA packaging. One of them is a 52-base-pair deletion encompassing the putative encapsidation signal in the leader region. The 3' long terminal repeat was also removed and replaced by the polyadenylation sequence from the herpes simplex virus thymidine kinase gene. When pHF-g cells were transfected by an avian leukosis virus-based vector, they produced replication-defective virus at high titer but they did not release any replication-competent particles. Proviral DNA was shown to be correctly integrated as well as correctly expressed. Viral RNAs were shown to be correctly translated into gag-related polypeptides.
Savatier
P
P
Laboratoire de Biologie Cellulaire, INRA, CNRS UA92, Université Claude Bernard Lyon I, Villeurbanne, France.
Bagnis
C
C
Thoraval
P
P
Poncet
D
D
Belakebi
M
M
Mallet
F
F
Legras
C
C
Cosset
F L
FL
Thomas
J L
JL
Chebloune
Y
Y
D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics*; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular / Q000379:methods*; D004279:DNA, Viral / Q000235:genetics; D003673:Defective Viruses / Q000235:genetics / Q000502:physiology; D005814:Genes, Viral; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics*; D012367:RNA, Viral / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication
D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins