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Updated: Aug 11, 2026
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Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
Published on: September 21, 2017
Initial mechanistic studies of antisense targeting in cells
Xinrong Liu1, Kayoko Nakamura, Yi Wang
1Department of Radiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
Summary
Antisense DNA effectively targets intracellular mdr1 mRNA, acting as a primer for complementary DNA elongation. High accumulation suggests targeting of nuclear pre-mRNA, not cytoplasmic mRNA, at high transcription rates.
Area of Science:
- Molecular Biology
- Antisense Technology
Background:
- Antisense targeting is a promising therapeutic strategy.
- Understanding the precise mechanism of intracellular antisense targeting is crucial for its continued development.
Purpose of the Study:
- To elucidate the mechanism of intracellular antisense targeting.
- To investigate the accumulation and localization of antisense DNA in cells.
- To determine the effect of antisense DNA on mRNA transcription rates.
Main Methods:
- In situ transcription, immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR) were employed.
- Nuclear accumulation of 99mTc-labeled antisense DNA was measured.
- Messenger RNA (mRNA) transcription rates were assessed using novel radiolabeled probes.
Main Results:
- Antisense DNA demonstrated complementary DNA elongation, confirming intracellular targeting of mdr1 mRNA.
- Higher accumulation of antisense DNA was observed in the nucleus compared to the cytoplasm.
- No significant upregulation of mRNA levels was detected, ruling out a feedback mechanism.
Conclusions:
- The findings support antisense DNA as an effective intracellular targeting mechanism.
- High antisense DNA accumulation may be explained by targeting nuclear pre-mRNA at high transcription rates.
- The study provides insights into the localization and mechanism of antisense DNA action.
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