Next Article in Journal
A Novel Brucine Gel Transdermal Delivery System Designed for Anti-Inflammatory and Analgesic Activities
Previous Article in Journal
Serum Concentrations of Angiopoietin-2 and Soluble fms-Like Tyrosine Kinase 1 (sFlt-1) Are Associated with Coagulopathy among Patients with Acute Pancreatitis
Article Menu
Issue 4 (April) cover image

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2017, 18(4), 754; doi:10.3390/ijms18040754

CRISPR-Cas9 Mediated Gene-Silencing of the Mutant Huntingtin Gene in an In Vitro Model of Huntington’s Disease

1,2
,
1,2
,
1,2,3,4,5
,
1,2,6
and
1,2,3,4,*
1
Field Neurosciences Institute laboratory for Restorative Neurology at Central Michigan University, Mt. Pleasant, MI 48859, USA
2
Program in Neuroscience, Central Michigan University, Mt. Pleasant, MI 48859, USA
3
Department of Psychology, Central Michigan University, Mt. Pleasant, MI 48859, USA
4
Field Neurosciences Institute, St. Mary’s of Michigan, Saginaw, MI 48604, USA
5
Department of Biology, Saginaw Valley State University, Saginaw, MI 48604, USA
6
College of Medicine, Central Michigan University, Mt. Pleasant, MI 48859, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Irmgard Tegeder
Received: 16 February 2017 / Revised: 23 March 2017 / Accepted: 26 March 2017 / Published: 2 April 2017
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
View Full-Text   |   Download PDF [2924 KB, uploaded 5 April 2017]   |  

Abstract

Huntington’s disease (HD) is a fatal neurodegenerative genetic disease characterized by a loss of neurons in the striatum. It is caused by a mutation in the Huntingtin gene (HTT) that codes for the protein huntingtin (HTT). The mutant Huntingtin gene (mHTT) contains extra poly-glutamine (CAG) repeats from which the translated mutant huntingtin proteins (mHTT) undergo inappropriate post-translational modifications, conferring a toxic gain of function, in addition to its non-functional property. In order to curb the production of the mHTT, we have constructed two CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR associate protein) plasmids, among which one nicks the DNA at untranslated region upstream to the open reading frame (uORF), and the other nicks the DNA at exon1-intron boundary. The primary goal of this study was to apply this plasmid into mesenchymal stem cells (MSCs) extracted from the bone-marrow of YAC128 mice, which carries the transgene for HD. Our results suggest that the disruption of uORF through CRISPR-Cas9 influences the translation of mHTT negatively and, to a lesser extent, disrupts the exon1-intron boundary, which affects the translation of the mHTT. These findings also revealed the pattern of the nucleotide addition or deletion at the site of the DNA-nick in this model. View Full-Text
Keywords: Huntington’s disease; CAG repeat; mutant huntingtin; gene editing; CRISPR-Cas9 system; pattern of NHEJ; YAC128; Kozak sequence Huntington’s disease; CAG repeat; mutant huntingtin; gene editing; CRISPR-Cas9 system; pattern of NHEJ; YAC128; Kozak sequence
Figures

Figure 1

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).

Supplementary material

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

Kolli, N.; Lu, M.; Maiti, P.; Rossignol, J.; Dunbar, G.L. CRISPR-Cas9 Mediated Gene-Silencing of the Mutant Huntingtin Gene in an In Vitro Model of Huntington’s Disease. Int. J. Mol. Sci. 2017, 18, 754.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

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