Högskolan i Skövde

his.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • apa-cv
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Epigenome targeting of immune genes involved in autoimmunity of multiple sclerosis
University of Skövde, School of Bioscience.
2025 (English)Independent thesis Basic level (degree of Bachelor), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Multiple sclerosis is the most common chronic inflammatory and neurodegenerative disease of the central nervous system, highly influenced by genetic factors in combination with the epigenetic and environmental factors. Among the strongest genetic risk factors is the DR15 haplotype, particularly the class II HLA-DRB1*15:01 allele, whose expression is highly regulated by the Class II translational activator, CIITA. This study aimed to investigate how targeted DNA methylation influences the expression of HLA class II genes, specifically focused on CIITA. THP-1 cells stably expressing dCas9-DNMT3a were treated with different pro-inflammatory stimuli, such as hIFN-γ and LPS, to determine under which conditions there is an increased expression of HLA class II genes in these cells. This resulted in the establishment of a baseline immune activation model, which was later used to assess how targeted DNA methylation at the CIITA promoter and HLA-DRB1*15:01 locus influences gene expression. Guide RNAs targeting promoter regions of CIITA, namely pIII and pIV, were delivered via lentiviral transduction, and the changes in gene expression and promoter methylation were assessed using RT-qPCR and pyrosequencing. The stimulation of THP-1 stable cells with hIFN-γ showed robustly induced expression of CIITA and HLA class II genes, while LPS alone showed no effect. The results of gRNA-mediated targeting showed minor changes in expression levels, and the pyrosequencing results revealed no consistent increase in DNA methylation at the targeted promoter regions. These findings highlight both the potential but also the current limitations of dCas9-DNMT3a-mediated epigenetic editing, with future improvements in gRNA design, delivery efficiency, and further research in target selection needed for more accurate results and better effect in transcriptional silencing. This study provides insights into epigenetic regulation of immune genes relevant to multiple sclerosis, but further investigation is needed.

Place, publisher, year, edition, pages
2025. , p. 34
National Category
Medical Bioscience
Identifiers
URN: urn:nbn:se:his:diva-25639OAI: oai:DiVA.org:his-25639DiVA, id: diva2:1985845
External cooperation
Center for Molecular Medicine, Department of Clinical Neuroscience, Karolinska Institutet
Subject / course
Bioscience
Educational program
Bioscience - Molecular Biodesign
Supervisors
Examiners
Available from: 2025-07-28 Created: 2025-07-28 Last updated: 2025-09-29Bibliographically approved

Open Access in DiVA

fulltext(1261 kB)121 downloads
File information
File name FULLTEXT01.pdfFile size 1261 kBChecksum SHA-512
974527c84064b94a17cb5ab4e6fefdd512cd661769942b47f74314d712dd2ec650e6ba40f23b64452257be123ad7f1f8cb5e90851d5a91f47bf3391a6169783e
Type fulltextMimetype application/pdf

By organisation
School of Bioscience
Medical Bioscience

Search outside of DiVA

GoogleGoogle Scholar
Total: 123 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

urn-nbn

Altmetric score

urn-nbn
Total: 248 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • apa-cv
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf