title: Antisense-mediated exon skipping to shift alternative splicing to treat cancer.
abstract: Manipulation of alternative splicing is a unique way of regulating gene expression. Here, a detailed protocol is presented for modification of HER2 pre-mRNA alternative splicing. A 20-mer splice switching oligonucleotide (SSO) targeting exon 15 in HER2 pre-mRNA induced skipping of exon 15, resulted in significant downregulation of full-length HER2 mRNA and protein expression in HER2-overexpressing breast cancer cell line SK-BR-3.
authors.affiliation: University of North Carolina, Chapel Hill, NC, USA. jingwan@med.unc.edu
authors.forename: Jing
authors.initials: J
authors.lastname: Wan
mesh_terms: D017398:Alternative Splicing; D015153:Blotting, Western / Q000379:methods; D001943:Breast Neoplasms / Q000235:genetics* / Q000628:therapy; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005091:Exons; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000627:therapeutic use*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012322:RNA Precursors / Q000235:genetics* / Q000302:isolation & purification; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D016376:Oligonucleotides, Antisense; D012313:RNA; D012322:RNA Precursors; D018719:Erb-b2 Receptor Tyrosine Kinases