Positional effects of the central DNA flap in HIV-1-derived lentiviral vectors.
During reverse transcription of the human immunodeficiency virus type 1 (HIV-1), a 90- or 99-nucleotide long DNA flap is formed at the centre of the viral cDNA. The presence of a central DNA flap in lentiviral vectors improves transduction efficiency significantly. We analysed the stimulation of lentiviral vector transduction by a DNA flap present at ectopic positions in the viral cDNA. A HIV-1 vector containing the cPPT/CTS fragment immediately downstream of the 5'-LTR performed as well as the wild-type cPPT-vector. Cloning of the cPPT/CTS fragment in front of the 3'-LTR resulted as well in a vector with higher transduction efficiency than a vector without central flap. These results demonstrate that the position of the DNA flap is not essential for its function in the context of HIV-1-derived lentiviral vectors. This may have consequences for vector design and our understanding of the functioning of the HIV-1 DNA flap.
De Rijck
Jan
J
Laboratory for Molecular Virology and Gene Therapy, KULeuven and KULAK, Flanders, Belgium.
Van Maele
Bénédicte
B
Debyser
Zeger
Z
D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004279:DNA, Viral / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000502:physiology*; D016086:Lentivirus / Q000235:genetics*; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D009690:Nucleic Acid Conformation; D013329:Structure-Activity Relationship; D014162:Transfection / Q000379:methods*
D004279:DNA, Viral