Downregulation of gene expression with negatively charged peptide nucleic acids (PNAs) in zebrafish embryos.
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.
Wickstrom
Eric
E
Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Urtishak
Karen A
KA
Choob
Michael
M
Tian
Xiaobing
X
Sternheim
Nitzan
N
Cross
Laura M
LM
Rubinstein
Amy
A
Farber
Steven A
SA
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
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