Regulation of in vitro gene expression using antisense oligonucleotides or antisense expression plasmids transfected using starburst PAMAM dendrimers.
Starburst polyamidoamine (PAMAM) dendrimers are a new type of synthetic polymer characterized by a branched spherical shape and a high density surface charge. We have investigated the ability of these dendrimers to function as an effective delivery system for antisense oligonucleotides and 'antisense expression plasmids' for the targeted modulation of gene expression. Dendrimers bind to various forms of nucleic acids on the basis of electrostatic interactions, and the ability of DNA-dendrimer complexes to transfer oligonucleotides and plasmid DNA to mediate antisense inhibition was assessed in an in vitro cell culture system. Cell lines that permanently express luciferase gene were developed using dendrimer mediated transfection. Transfections of antisense oligonucleotides or antisense cDNA plasmids into these cell lines using dendrimers resulted in a specific and dose dependent inhibition of luciferase expression. This inhibition caused approximately 25-50% reduction of baseline luciferase activity. Binding of the phosphodiester oligonucleotides to dendrimers also extended their intracellular survival. While dendrimers were not cytotoxic at the concentrations effective for DNA transfer, some non-specific suppression of luciferase expression was observed. Our results indicate that Starburst dendrimers can be effective carriers for the introduction of regulatory nucleic acids and facilitate the suppression of the specific gene expression.
Bielinska
A
A
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109-0666, USA.
Kukowska-Latallo
J F
JF
Johnson
J
J
Tomalia
D A
DA
Baker
J R
JR
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary; D004337:Drug Carriers; D004355:Drug Stability; D004563:Electrochemistry; D005786:Gene Expression Regulation; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D010957:Plasmids; D011108:Polymers / Q000737:chemistry; D051381:Rats; D014162:Transfection
D016373:DNA, Antisense; D018076:DNA, Complementary; D004337:Drug Carriers; D016376:Oligonucleotides, Antisense; D011108:Polymers; D008156:Luciferases