title: Downregulation of gene expression with negatively charged peptide nucleic acids (PNAs) in zebrafish embryos.
abstract: We found that negatively charged, highly soluble PNA analogs with alternating phosphonates (HypNA-pPNAs) are effective and specific antisense agents in zebrafish embryos, showing comparable potency and greater specificity against chordin, ntl and uroD. In addition, we successfully phenocopied a dharma mutant that had not been found susceptible to MO knockdown. Both MO and HypNA-pPNAs against a tumor suppressor gene induced comparable upregulation of p53, illustrating similar effects on transcription profiles. HypNA-pPNAs are therefore a valuable alternative for reverse genetic studies, enabling the targeting of previously inaccessible genes in zebrafish or validating newly identified orthologs, and perhaps for reverse genetic studies in other organisms.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Wickstrom
authors.forename: Karen A
authors.initials: KA
authors.lastname: Urtishak
authors.forename: Michael
authors.initials: M
authors.lastname: Choob
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Tian
authors.forename: Nitzan
authors.initials: N
authors.lastname: Sternheim
authors.forename: Laura M
authors.initials: LM
authors.lastname: Cross
authors.forename: Amy
authors.initials: A
authors.lastname: Rubinstein
authors.forename: Steven A
authors.initials: SA
authors.lastname: Farber
mesh_terms: D000818:Animals; D000838:Anions / Q000737:chemistry / Q000494:pharmacology; D015536:Down-Regulation; D004625:Embryo, Nonmammalian / Q000187:drug effects; D005326:Fetal Proteins; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006023:Glycoproteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000494:pharmacology*; D020825:T-Box Domain Proteins / Q000235:genetics; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics; D000098908:Brachyury Protein
chemical_list: D000838:Anions; D005326:Fetal Proteins; D006023:Glycoproteins; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; D016376:Oligonucleotides, Antisense; D020135:Peptide Nucleic Acids; D020825:T-Box Domain Proteins; D029961:Zebrafish Proteins; D000098908:Brachyury Protein; C113874:dharma protein, zebrafish; C054234:chordin