Use of triplex-forming oligonucleotides and adenoviral constructs for studying the regulation of gene expression.
Short synthetic homopyrimidine- or homopurine-rich oligonucleotides can form sequence-specific triplexes with corresponding homopurine-homopyrimidine sites on duplex DNA and block transcription of a target gene in vitro. Such triplex-forming oligonucleotides (TFOs) can be rationally designed to target homopurine/homopyrimidine sequences that are often found in eukaryotic genes and thus used to modulate the expression of these genes. The antigene strategy using TFOs has been successfully applied to a number of genes in vitro. In this article we describe methods used in applying this antigene approach to the rat aquaporin 5 (rAQP5) gene. We specifically focus on the selection of TFOs based on the sequence of the target gene and on a novel method employing adenoviruses for delivery of TFOs to cells in vitro.
Shamsul Hoque
A T
AT
National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
Panyutin
I G
IG
Baum
B J
BJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D051402:Aquaporin 5; D020346:Aquaporins / Q000235:genetics*; D002460:Cell Line; D004247:DNA / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004285:Dogs; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005786:Gene Expression Regulation / Q000235:genetics*; D017930:Genes, Reporter; D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D051381:Rats; D014162:Transfection
C496040:Aqp5 protein, rat; D051402:Aquaporin 5; D020346:Aquaporins; D004279:DNA, Viral; D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides; C051802:triplex DNA; D011107:Polylysine; D011094:Polyethyleneimine; D004247:DNA