title: In vivo site-directed mutagenesis of the factor IX gene by chimeric RNA/DNA oligonucleotides.
abstract: A chimeric RNA/DNA oligonucleotide was constructed to induce a sequence mutation in the rat factor IX gene, resulting in prolonged coagulation. Oligonucleotides were targeted to hepatocytes in cell culture or in vivo by intravenous injection. Nucleotide conversion was both site-specific and dose-dependent. The mutated gene was associated in vivo with significantly reduced factor IX coagulant activity and a marked prolongation of the activated partial thromboplastin time. The results demonstrate that single base-pair alterations can be introduced in hepatocytes in situ by RNA/DNA oligonucleotides, suggesting a potentially powerful strategy for hepatic gene repair without the use of viral vectors.
authors.affiliation: Department of Medicine, University of Minnesota Medical School, Minneapolis 55455, USA.
authors.forename: B T
authors.initials: BT
authors.lastname: Kren
authors.forename: P
authors.initials: P
authors.lastname: Bandyopadhyay
authors.forename: C J
authors.initials: CJ
authors.lastname: Steer
mesh_terms: D000818:Animals; D001692:Biological Transport; D002469:Cell Separation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005164:Factor IX / Q000032:analysis / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D008297:Male; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D009843:Oligoribonucleotides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017422:Sequence Analysis, DNA; D012694:Serine / Q000235:genetics; D014162:Transfection
chemical_list: D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D012694:Serine; D005164:Factor IX