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Issue 8, 2017
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RNA splicing process analysis for identifying antisense oligonucleotide inhibitors with padlock probe-based isothermal amplification

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Abstract

RNA splicing, which mainly involves two transesterification steps, is a fundamental process of gene expression and its abnormal regulation contributes to serious genetic diseases. Antisense oligonucleotides (ASOs) are genetic control tools that can be used to specifically control genes through alteration of the RNA splicing pathway. Despite intensive research, how ASOs or various other factors influence the multiple processes of RNA splicing still remains obscure. This is largely due to an inability to analyze the splicing efficiency of each step in the RNA splicing process with high sensitivity. We addressed this limitation by introducing a padlock probe-based isothermal amplification assay to achieve quantification of the specific products in different splicing steps. With this amplified assay, the roles that ASOs play in RNA splicing inhibition in the first and second steps could be distinguished. We identified that 5′-ASO could block RNA splicing by inhibiting the first step, while 3′-ASO could block RNA splicing by inhibiting the second step. This method provides a versatile tool for assisting efficient ASO design and discovering new splicing modulators and therapeutic drugs.

Graphical abstract: RNA splicing process analysis for identifying antisense oligonucleotide inhibitors with padlock probe-based isothermal amplification

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Supplementary files

Article information


Submitted
25 Mar 2017
Accepted
07 Jun 2017
First published
13 Jun 2017

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2017,8, 5692-5698
Article type
Edge Article

RNA splicing process analysis for identifying antisense oligonucleotide inhibitors with padlock probe-based isothermal amplification

X. Ren, R. Deng, L. Wang, K. Zhang and J. Li, Chem. Sci., 2017, 8, 5692 DOI: 10.1039/C7SC01336A

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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