title: Correction of chromosomal point mutations in human cells with bifunctional oligonucleotides.
abstract: A sequence-specific genomic delivery system for the correction of chromosomal mutations was designed by incorporating two different binding domains into a single-stranded oligonucleotide. A repair domain (RD) contained the native sequence of the target region. A third strand-forming domain (TFD) was designed to form a triplex by Hoogsteen interactions. The design was based upon the premise that the RD will rapidly form a heteroduplex that is anchored synergistically by the TFD. Deoxyoligonucleotides were designed to form triplexes in the human adenosine deaminase (ADA) and p53 genes adjacent to known point mutations. Transfection of ADA-deficient human lymphocytes corrected the mutant sequence in 1-2% of cells. Neither the RD or TFD individually corrected the mutation. Transfection of p53 mutant human glioblastoma cells corrected the mutation and induced apoptosis in 7.5% of cells.
authors.affiliation: Novartis Pharmaceuticals Corp., East Hanover, NJ, 07936, USA. ken.culver@pharma.novartis.com
authors.forename: K W
authors.initials: KW
authors.lastname: Culver
authors.forename: W T
authors.initials: WT
authors.lastname: Hsieh
authors.forename: Y
authors.initials: Y
authors.lastname: Huyen
authors.forename: V
authors.initials: V
authors.lastname: Chen
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.forename: Y
authors.initials: Y
authors.lastname: Khripine
authors.forename: A
authors.initials: A
authors.lastname: Khorlin
mesh_terms: D000243:Adenosine Deaminase / Q000235:genetics*; D001483:Base Sequence; D002461:Cell Line, Transformed; D002877:Chromosomes, Human; D016158:Genes, p53 / Q000235:genetics; D006801:Humans; D009841:Oligonucleotides / Q000494:pharmacology*; D017354:Point Mutation
chemical_list: D009841:Oligonucleotides; D000243:Adenosine Deaminase