Date of Award
Spring 2026
Language
English
Embargo Period
4-30-2028
Document Type
Master's Thesis
Degree Name
Master of Science (MS)
College/School/Department
Department of Biological Sciences
Program
Biology
First Advisor
Dr. Kaalak Reddy
Committee Members
Dr. J. Andrew Berglund, Dr. Mark Handley, Dr. Gabriele Fuchs
Subject Categories
Biology
Abstract
Myotonic dystrophy is the most common adult-onset muscular dystrophy and exists in two genetically different forms: myotonic dystrophy type 1 (DM1), caused by CTG repeat expansions in the DMPK gene and myotonic dystrophy type 2 (DM2) caused by CCTG repeat expansions in the CNBP gene. Despite sharing similar pathogenic mechanisms with DM1, DM2 remains comparatively underexplored, in part due to the limited availability of robust and controllable cellular models. To address this gap, this study aimed to develop and validate an inducible cellular system for expression of expanded CCUG repeat RNAtogether with control RNA. Expression cassettes containing control and expanded CCTG repeat sequences were designed with a tetracycline-responsive promoter and cloned into a piggyBac transposon vector backbone for stable genomic integration. The constructs included a 6-stop sequence to prevent unintended translation and a 3×3 tag to facilitate detection of potential repeat-associated non-AUG (RAN) translation products. Using a control cassette as the starting template, we inserted a new probe-binding sequence to enable later distinction of the CCTG cassette from the control construct. Polyadenylation signal was included to ensure proper transcript termination and stability. Stable cell lines were generated in HeLa and HEK293 cells via transposase-mediated integration, followed by antibiotic selection and clonal isolation by FACS sorting. Genomic integration and relative copy number across clones were assessed using PCR. Functional validation of selected clones demonstrated that doxycycline treatment induced expression of repeat-containing transcripts. This study establishes multiple DM2 model cell lines. These could provide a foundation for future mechanistic and therapeutic screening.
License
This work is licensed under the University at Albany Standard Author Agreement.
Recommended Citation
Devpura, Damini, "Development of an Inducible Cellular Model for Myotonic Dystrophy Type 2 (DM2)" (2026). Electronic Theses & Dissertations (2024 - present). 484.
https://scholarsarchive.library.albany.edu/etd/484