An intracellular kinase signal-responsive gene carrier for disordered cell-specific gene therapy.
We have previously reported artificial gene-regulation systems responding to cyclic AMP-dependent protein kinase (PKA) using cationic polymer. This cationic polymer (PAK) was a graft-type polymer with an oligopeptide that is a substrate for PKA and could regulate gene-expression in a cell-free system. In the present study, we carried out a detailed characterization of the PAK-DNA complex (AFM observation and DLS measurement) and tried to apply this polymer to living cells. In the unstimulated NIH 3T3 cells, transfection of the PAK-DNA complex showed no expression of the delivered gene. This means that PAK formed a stable complex with DNA in the normal cells to totally suppress gene expression. In contrast, significant expression was seen when the PAK-DNA complex was delivered to forskolin-treated cells. Thus, activated PKA disintegrates the complexes even in living cells, resulting in gene expression. Our results indicate that this type of intracellular signal-responsive polymer will be useful for the cell-specific release of genes.
Oishi
Jun
J
Graduate School of Systems Life Sciences, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan; CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan.
Kawamura
Kenji
K
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan; CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan.
Kang
Jeong-Hun
JH
CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan.
Kodama
Kota
K
CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan.
Sonoda
Tatsuhiko
T
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan.
Murata
Masaharu
M
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan; CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan.
Niidome
Takuro
T
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan; CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan.
Katayama
Yoshiki
Y
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan; CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi-shi, Saitama 332-0012, Japan. Electronic address: ykatatcm@mbox.nc.kyushu-u.ac.jp.
D016475:3T3 Cells; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004337:Drug Carriers; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008027:Light; D008156:Luciferases / Q000235:genetics; D051379:Mice; D018625:Microscopy, Atomic Force; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011108:Polymers / Q000138:chemical synthesis; D012542:Scattering, Radiation; D013778:Tetrazolium Salts / Q000737:chemistry
C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D004337:Drug Carriers; D010455:Peptides; D011108:Polymers; D013778:Tetrazolium Salts; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate; D004247:DNA; D008156:Luciferases; D017868:Cyclic AMP-Dependent Protein Kinases