Quantification of oxidized levels of specific RNA species using an aldehyde reactive probe

Tanaka, Mikiei; Han, Song; Küpfer, Pascal A.; Leumann, Christian J.; Sonntag, William E. (2011). Quantification of oxidized levels of specific RNA species using an aldehyde reactive probe. Analytical biochemistry, 417(1), pp. 142-148. San Diego, Calif.: Elsevier 10.1016/j.ab.2011.05.038

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Emerging evidence has shown that oxidation of RNA, including messenger RNA (mRNA), is elevated in several age-related diseases, although investigation of oxidized levels of individual RNA species has been limited. Recently we reported that an aldehyde reactive probe (ARP) quantitatively reacts with oxidatively modified depurinated/depyrimidinated (abasic) RNA. Here we report a novel method to isolate oxidized RNA using ARP and streptavidin beads. An oligo RNA containing abasic sites that were derivatized with ARP was pulled down by streptavidin beads, whereas a control oligo RNA was not. In vitro oxidized RNA, as well as total cellular RNA, isolated from oxidatively stressed cells was also pulled down, dependent on oxidation level, and concentrated in the pull-down fraction. Quantitative reverse transcription polymerase chain reaction (RT-PCR) using RNA in the pull-down fraction demonstrated that several gene transcripts were uniquely increased in the fraction by oxidative stress. Thus, our method selectively concentrates oxidized RNA by pull-down and enables the assessment of oxidation levels of individual RNA species. (C) 2011 Elsevier Inc. All rights reserved.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Küpfer, Pascal and Leumann, Christian

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

0003-2697

Publisher:

Elsevier

Language:

English

Submitter:

Christian Leumann

Date Deposited:

04 Oct 2013 14:26

Last Modified:

10 Aug 2015 11:25

Publisher DOI:

10.1016/j.ab.2011.05.038

Web of Science ID:

000293548400018

BORIS DOI:

10.7892/boris.9418

URI:

https://boris.unibe.ch/id/eprint/9418 (FactScience: 215152)

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