Reliable Resolution of Full-Length Huntingtin Alleles by Quantitative Immunoblotting.

Journal of Huntington's Disease
N. S. CaronM. R. Hayden

Abstract

Therapeutics that lower mutant huntingtin (mHTT) have shown promise in preclinical studies and are in clinical development for the treatment of Huntington disease (HD). Multiple assays have been developed that either quantify mHTT or total HTT but may not accurately measure levels of wild type HTT (wtHTT) in biological samples. To optimize a method that can be used to resolve, quantify and directly compare levels of full length wtHTT and mHTT in HD samples. We provide a detailed quantitative immunoblotting protocol to reproducibly resolve full length wtHTT and mHTT in multiple HD mouse and patient samples. We show that this assay can be modified, depending on the sample, to resolve wtHTT and mHTT with a wide range of polyglutamine length differences (ΔQs 22-179). We also demonstrate that this method can be used to quantify allele-selective lowering of mHTT using an antisense oligonucleotide in HD patient-derived cells. This quantitative immunoblotting method can be used to reliably resolve full-length HTT alleles with ΔQs≥22 and allows for direct comparison of wtHTT and mHTT levels in HD samples.

References

Apr 26, 1995·Biochemical and Biophysical Research Communications·K IdeI Kanazawa
Jun 1, 1996·Neurobiology of Disease·F PersichettiM E MacDonald
Jun 19, 2003·Human Molecular Genetics·Elizabeth J SlowMichael R Hayden
Aug 20, 2003·The Journal of Comparative Neurology·Liliana B MenalledMarie-Françoise Chesselet
Sep 2, 2004·Molecular and Cellular Biology·Eugenia TrushinaCynthia T McMurray
Jun 14, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Michelle GrayX William Yang
Dec 20, 2008·Journal of Neurochemistry·Yu ZhangRobert M Friedlander
Feb 26, 2009·Molecular Therapy : the Journal of the American Society of Gene Therapy·Ryan L BoudreauBeverly L Davidson
Jul 29, 2010·Physiological Reviews·Chiara ZuccatoElena Cattaneo
Oct 6, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jeffrey B CarrollMichael R Hayden
Oct 28, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jodi L McBrideBeverly L Davidson
Mar 1, 2012·The Journal of Biological Chemistry·Ellen SappMarian DiFiglia
Sep 22, 2012·The Journal of Clinical Investigation·Andreas WeissSarah J Tabrizi
Sep 25, 2012·Human Molecular Genetics·Amber L SouthwellMichael R Hayden
Oct 12, 2012·Proceedings of the National Academy of Sciences of the United States of America·Mireia Garriga-CanutMark Isalan
May 13, 2014·PloS One·Douglas MacdonaldIgnacio Munoz-Sanjuan
Aug 8, 2014·Molecular Therapy : the Journal of the American Society of Gene Therapy·Amber L SouthwellMichael R Hayden
Apr 7, 2015·The Journal of Clinical Investigation·Edward J WildAndreas Weiss
Jul 24, 2015·Molecular Therapy : the Journal of the American Society of Gene Therapy·Chris KayMichael R Hayden
Apr 25, 2017·Scientific Reports·James R C MillerSarah J Tabrizi
Mar 8, 2018·Acta Neuropathologica Communications·Dagmar E EhrnhoeferMichael R Hayden
Oct 5, 2018·Science Translational Medicine·Amber L SouthwellMichael R Hayden
Dec 31, 2018·Journal of Huntington's Disease·Adelaide TousleyKimberly B Kegel-Gleason

❮ Previous
Next ❯

Citations

Aug 15, 2021·Neurobiology of Disease·Fanny L LemariéMichael R Hayden

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antisense Oligonucleotides: ND

This feed focuses on antisense oligonucleotide therapies such as Inotersen, Nusinursen, and Patisiran, in neurodegenerative diseases including amyotrophic lateral sclerosis.

Related Papers

Frontiers in Cellular Neuroscience
Jessica C BarronMatthew P Parsons
BioRxiv : the Preprint Server for Biology
Daniele BertoglioI. Munoz-Sanjuan
The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
Nicholas S CaronAmber L Southwell
© 2021 Meta ULC. All rights reserved