Target Gene Knockdown by 2',4'-BNA/LNA Antisense Oligonucleotides in Zebrafish
- Authors
- Itoh, M., Nakaura, M., Imanishi, T., and Obika, S.
- ID
- ZDB-PUB-140321-37
- Date
- 2014
- Source
- Nucleic Acid Therapeutics 24(3): 186-91 (Journal)
- Registered Authors
- Itoh, Motoyuki
- Keywords
- none
- MeSH Terms
-
- Animals
- Base Sequence
- Codon, Initiator
- Embryo, Nonmammalian
- Gene Expression Regulation, Developmental
- Gene Knockdown Techniques*
- Molecular Sequence Data
- Morpholinos/genetics
- Morpholinos/metabolism
- Oligonucleotides/chemical synthesis
- Oligonucleotides/genetics*
- Oligonucleotides/metabolism
- Oligonucleotides, Antisense/chemical synthesis
- Oligonucleotides, Antisense/genetics*
- Oligonucleotides, Antisense/metabolism
- Protein Biosynthesis
- Ribonuclease H/metabolism
- Transcription Factor 7-Like 1 Protein/antagonists & inhibitors
- Transcription Factor 7-Like 1 Protein/genetics*
- Transcription Factor 7-Like 1 Protein/metabolism
- Tumor Suppressor Protein p53/genetics
- Tumor Suppressor Protein p53/metabolism
- Zebrafish
- Zebrafish Proteins/antagonists & inhibitors
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- PubMed
- 24460393 Full text @ Nucleic Acid Ther.
Gene knockdowns using oligonucleotide-based approaches are useful for studying gene function in both in vitro cell culture systems and in vivo animal models. We evaluated the efficacy of 22,42-bridged nucleic acids (BNA)-modified antisense oligonucleotides (AONs) for gene knockdown in zebrafish. We used the tcf7l1a gene as a model for testing the knockdown efficacy of 22,42-BNA AONs and examined how the target sites/affinity and RNase H induction activity of 22,42-BNA AONs affect knockdown efficacy. We found that tcf7l1a gene function was knocked down by 22,42-BNA AONs that target the start codon and induce RNase H activity. Although nonspecific p53-mediated developmental defects were observed at higher doses, the effective dose of the 22,42-BNA AONs for tcf7l1a is much lower than that of morpholino oligonucleotides. Our data thus show a potential application for 22,42-BNA AONs in the downregulation of specific genes in zebrafish.