Splice-switching efficiency and specificity for oligonucleotides with locked nucleic acid monomers

2008 ◽  
Vol 412 (2) ◽  
pp. 307-313 ◽  
Author(s):  
Peter Guterstam ◽  
Maria Lindgren ◽  
Henrik Johansson ◽  
Ulf Tedebark ◽  
Jesper Wengel ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
High Efficiency ◽  
Locked Nucleic Acid ◽  
Switching Activity ◽  
Mismatch Discrimination ◽  
Therapeutic Platform ◽  
Splicing Patterns ◽  
Switching Efficiency ◽  
Target Effects

The use of antisense oligonucleotides to modulate splicing patterns has gained increasing attention as a therapeutic platform and, hence, the mechanisms of splice-switching oligonucleotides are of interest. Cells expressing luciferase pre-mRNA interrupted by an aberrantly spliced β-globin intron, HeLa pLuc705, were used to monitor the splice-switching activity of modified oligonucleotides by detection of the expression of functional luciferase. It was observed that phosphorothioate 2′-O-methyl RNA oligonucleotides containing locked nucleic acid monomers provide outstanding splice-switching activity. However, similar oligonucleotides with several mismatches do not impede splice-switching activity which indicates a risk for off-target effects. The splice-switching activity is abolished when mismatches are introduced at several positions with locked nucleic acid monomers suggesting that it is the locked nucleic acid monomers that give rise to low mismatch discrimination to target pre-mRNA. The results highlight the importance of rational sequence design to allow for high efficiency with simultaneous high mismatch discrimination for splice-switching oligonucleotides and suggest that splice-switching activity is tunable by utilizing locked nucleic acid monomers.

scholarly journals Modulating Anti-MicroRNA-21 Activity and Specificity Using Oligonucleotide Derivatives and Length Optimization

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Andrés Muñoz-Alarcón ◽  
Peter Guterstam ◽  
Cristian Romero ◽  
Mark A. Behlke ◽  
Kim A. Lennox ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
Posttranscriptional Regulation ◽  
Regulation Of Gene Expression ◽  
Locked Nucleic Acid ◽  
Cellular Functions ◽  
Mismatch Discrimination ◽  
Increased Activity ◽  
Length Optimization ◽  
Oligonucleotide Derivatives

MicroRNAs are short, endogenous RNAs that direct posttranscriptional regulation of gene expression vital for many developmental and cellular functions. Implicated in the pathogenesis of several human diseases, this group of RNAs provides interesting targets for therapeutic intervention. Anti-microRNA oligonucleotides constitute a class of synthetic antisense oligonucleotides used to interfere with microRNAs. In this study, we investigate the effects of chemical modifications and truncations on activity and specificity of anti-microRNA oligonucleotides targeting microRNA-21. We observed an increased activity but reduced specificity when incorporating locked nucleic acid monomers, whereas the opposite was observed when introducing unlocked nucleic acid monomers. Our data suggest that phosphorothioate anti-microRNA oligonucleotides yield a greater activity than their phosphodiester counterparts and that a moderate truncation of the anti-microRNA oligonucleotide improves specificity without significantly losing activity. These results provide useful insights for design of anti-microRNA oligonucleotides to achieve both high activity as well as efficient mismatch discrimination.

scholarly journals Locked nucleic acid containing antisense oligonucleotides enhance inhibition of HIV-1 genome dimerization and inhibit virus replication

FEBS Letters ◽  
2004 ◽  
Vol 578 (3) ◽  
pp. 285-290 ◽  
Author(s):  
Joacim Elmén ◽  
Hong-Yan Zhang ◽  
Bartek Zuber ◽  
Karl Ljungberg ◽  
Britta Wahren ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Virus Replication ◽  
Antisense Oligonucleotides ◽  
Locked Nucleic Acid ◽  
Inhibit Virus Replication ◽  
Genome Dimerization ◽  
Hiv 1

scholarly journals An Investigation into the Potential of Targeting Escherichia coli rne mRNA with Locked Nucleic Acid (LNA) Gapmers as an Antibacterial Strategy

Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3414
Author(s):  
Layla R. Goddard ◽  
Charlotte E. Mardle ◽  
Hassan Gneid ◽  
Ciara G. Ball ◽  
Darren M. Gowers ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Nucleic Acid ◽  
Translation Initiation ◽  
Antisense Oligonucleotide ◽  
Antisense Oligonucleotides ◽  
Mrna Translation ◽  
Rna Degradation ◽  
Rnase H ◽  
Locked Nucleic Acid ◽  
Translation Initiation Region

The increase in antibacterial resistance is a serious challenge for both the health and defence sectors and there is a need for both novel antibacterial targets and antibacterial strategies. RNA degradation and ribonucleases, such as the essential endoribonuclease RNase E, encoded by the rne gene, are emerging as potential antibacterial targets while antisense oligonucleotides may provide alternative antibacterial strategies. As rne mRNA has not been previously targeted using an antisense approach, we decided to explore using antisense oligonucleotides to target the translation initiation region of the Escherichia coli rne mRNA. Antisense oligonucleotides were rationally designed and were synthesised as locked nucleic acid (LNA) gapmers to enable inhibition of rne mRNA translation through two mechanisms. Either LNA gapmer binding could sterically block translation and/or LNA gapmer binding could facilitate RNase H-mediated cleavage of the rne mRNA. This may prove to be an advantage over the majority of previous antibacterial antisense oligonucleotide approaches which used oligonucleotide chemistries that restrict the mode-of-action of the antisense oligonucleotide to steric blocking of translation. Using an electrophoretic mobility shift assay, we demonstrate that the LNA gapmers bind to the translation initiation region of E. coli rne mRNA. We then use a cell-free transcription translation reporter assay to show that this binding is capable of inhibiting translation. Finally, in an in vitro RNase H cleavage assay, the LNA gapmers facilitate RNase H-mediated mRNA cleavage. Although the challenges of antisense oligonucleotide delivery remain to be addressed, overall, this work lays the foundations for the development of a novel antibacterial strategy targeting rne mRNA with antisense oligonucleotides.

scholarly journals Antisense oligonucleotides containing locked nucleic acid improve potency but cause significant hepatotoxicity in animals

2006 ◽  
Vol 35 (2) ◽  
pp. 687-700 ◽  
Author(s):  
Eric E. Swayze ◽  
Andrew M. Siwkowski ◽  
Edward V. Wancewicz ◽  
Michael T. Migawa ◽  
Tadeusz K. Wyrzykiewicz ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
Locked Nucleic Acid

scholarly journals Down-modulation of cancer targets using locked nucleic acid (LNA)-based antisense oligonucleotides without transfection

Gene Therapy ◽  
2010 ◽  
Vol 18 (4) ◽  
pp. 326-333 ◽  
Author(s):  
Y Zhang ◽  
Z Qu ◽  
S Kim ◽  
V Shi ◽  
B Liao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
Locked Nucleic Acid
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