Specific transgene expression in HIV-infected cells using protease-cleavable transcription regulator.
Gene therapy is a promising strategy for the treatment of HIV infection, but cell specificity remains an issue. Recently we have developed a new concept for a drug or gene delivery system responding to cellular signals (D-RECS) to achieve cell-specific transgene expression using a non-viral polymer-based vehicle. According to this concept, intracellular signaling enzymes, which are activated specifically in target cells, are used to trigger transgene expression. We previously applied this concept to HIV-1 protease and showed that the recombinant protease could act as a suitable signal. Here we further developed this system to achieve highly specific transgene expression in HIV-infected cells. We prepared a polymeric gene regulator grafted with a cationic peptide containing the HIV-Tat peptide via a specific substrate for HIV-1 protease. The regulator formed a stable polyplex with the transgene, suppressing its transcription. HIV-1 protease cleaved the peptide and released the transgene, which was consequently expressed specifically in activated HIV-infected cells, but remained unreleased and inactive in uninfected cells. The validity of this approach was further confirmed by applying it to the CVB1 2A protease of coxsackievirus (Picornaviridae family). This strategy should be widely applicable for specific expression of a variety of therapeutic genes in virus-infected cells.
Asai
Daisuke
D
Department of Microbiology, St. Marianna University School of Medicine, Kawasaki 216-8511, Japan. asai@marianna-u.ac.jp
Kuramoto
Masanori
M
Shoji
Yoko
Y
Kang
Jeong-Hun
JH
Kodama
Kota Bae
KB
Kawamura
Kenji
K
Mori
Takeshi
T
Miyoshi
Hiroshi
H
Niidome
Takuro
T
Nakashima
Hideki
H
Katayama
Yoshiki
Y
D002412:Cations / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry*; D015696:Gene Products, tat / Q000737:chemistry / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D015658:HIV Infections / Q000628:therapy; D016333:HIV Protease / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D019169:Jurkat Cells; D008156:Luciferases / Q000235:genetics; D010316:Particle Size; D010455:Peptides / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D014158:Transcription, Genetic; D019076:Transgenes
D002412:Cations; D004337:Drug Carriers; D015696:Gene Products, tat; D010455:Peptides; D011108:Polymers; D004247:DNA; D008156:Luciferases; D016333:HIV Protease; C514803:p16 protease, Human immunodeficiency virus 1