Next Article in Journal
Comparative Analysis of the Brassica napus Root and Leaf Transcript Profiling in Response to Drought Stress
Previous Article in Journal
LincRNA-p21: Implications in Human Diseases
Article Menu
Issue 8 (August) cover image

Export Article

Open AccessTechnical Note
Int. J. Mol. Sci. 2015, 16(8), 18741-18751; doi:10.3390/ijms160818741

In Vitro Expansion of CAG, CAA, and Mixed CAG/CAA Repeats

Department of Molecular Biomedicine, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14 Str., 61-704 Poznan, Poland
*
Author to whom correspondence should be addressed.
Academic Editor: Mateus Webba da Silva
Received: 27 July 2015 / Revised: 3 August 2015 / Accepted: 4 August 2015 / Published: 11 August 2015
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
View Full-Text   |   Download PDF [1681 KB, uploaded 11 August 2015]   |  

Abstract

Polyglutamine diseases, including Huntington’s disease and a number of spinocerebellar ataxias, are caused by expanded CAG repeats that are located in translated sequences of individual, functionally-unrelated genes. Only mutant proteins containing polyglutamine expansions have long been thought to be pathogenic, but recent evidence has implicated mutant transcripts containing long CAG repeats in pathogenic processes. The presence of two pathogenic factors prompted us to attempt to distinguish the effects triggered by mutant protein from those caused by mutant RNA in cellular models of polyglutamine diseases. We used the SLIP (Synthesis of Long Iterative Polynucleotide) method to generate plasmids expressing long CAG repeats (forming a hairpin structure), CAA-interrupted CAG repeats (forming multiple unstable hairpins) or pure CAA repeats (not forming any secondary structure). We successfully modified the original SLIP protocol to generate repeats of desired length starting from constructs containing short repeat tracts. We demonstrated that the SLIP method is a time- and cost-effective approach to manipulate the lengths of expanded repeat sequences. View Full-Text
Keywords: SLIP; trinucleotide repeats; in vitro expansion; repeats in vitro cloning; polyglutamine diseases; plasmid constructs SLIP; trinucleotide repeats; in vitro expansion; repeats in vitro cloning; polyglutamine diseases; plasmid constructs
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Figura, G.; Koscianska, E.; Krzyzosiak, W.J. In Vitro Expansion of CAG, CAA, and Mixed CAG/CAA Repeats. Int. J. Mol. Sci. 2015, 16, 18741-18751.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top