Investigation of requirements for efficient gene delivery using the HIV-1 based lentiviral transduction system.
The specific recognition and selection of the HIV-1 packaging signal psi (Psi) sequence which is mediated by Gag protein is believed to be pivotal for selective viral genomic RNA packaging and has been a basis for the development of HIV-based transgene delivery systems. However, the requirement of the psi sequence has been questioned recently by a report postulating that the psi element is not absolutely required for transgene transduction. Here, we used a four-plasmid transgene delivery system and analyzed the results by HIV p24 antigen assay, MT4 infection assay, HT1080 colony assay, and reverse transcription-PCR (RT-PCR). The results clearly demonstrate that the psi sequence must be present for efficient transgene encapsidation and transduction.
Kim
Seon Hee
SH
Department of Pathology, National Research Laboratory of Molecular Virology, School of Medicine, The Catholic University of Korea, Seocho-gu Seoul, Republic of Korea.
Jang
Soo In
SI
Park
Chul Yong
CY
You
Ji Chang
JC
D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D019065:Virus Assembly / Q000235:genetics*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins