Assessing average somatic CAG repeat instability at the protein level

0518791 - ÚŽFG 2020 RIV GB eng J - Journal Article
Aviolat, H. - Mouro Pinto, R. - Godshall, E. - Murtha, R. - Richey, H. E. - Sapp, E. - Vodička, Petr - Wheeler, W. C. - Kegel-Gleason, K. B. - DiFiglia, M.
Assessing average somatic CAG repeat instability at the protein level.
Scientific Reports. Roč. 9, DEC 16 (2019), č. článku 19152. ISSN 2045-2322. E-ISSN 2045-2322
Institutional support: RVO:67985904
Keywords : mutant huntingtin * CAG
OECD category: Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
Impact factor: 3.998, year: 2019
Method of publishing: Open access
https://www.nature.com/articles/s41598-019-55202-x

Sandwich ELISA-based methods use Abs that target the expanded polyglutamine (polyQ) tract to quantify mutant huntingtin (mHTT). Using Meso Scale Discovery (MSD) assay, the mHTT signal detected with MW1Ab correlated with polyQ length and doubled with a difference of only 7 glutamine residues between equivalent amounts of purified mHTTexon1 proteins. Similar polyQ length-dependent effects on MSD signals were confirmed using endogenous full length mHTT from brains of Huntington's disease (HD) knock-in (KI) mice. We used this avidity bias to devise a method to assess average CAG repeat instability at the protein level in a mixed population of HTT proteins present in tissues. Signal detected for average polyQ length quantification at the protein level by our method exhibited a strong correlation with average CAG repeat length at the genomic DNA level determined by PCR method in striatal tissue homogenates from Hdh(Q140) KI mice and in human HD postmortem cortex. This work establishes that CAG repeat instability in mutant HTT is reflected at the protein level.
Permanent Link: http://hdl.handle.net/11104/0303847