Date of Completion
2023
Document Type
Honors College Thesis
Department
Biochemistry
Thesis Type
College of Arts and Science Honors, Honors College
First Advisor
Jane B. Lian
Second Advisor
Matthias Brewer
Third Advisor
Lisa Schnell
Keywords
Osteogenesis, Osteoblast, LncRNA, Epigenetic Regulation, Gene Expression
Abstract
Long non-coding RNAs (lncRNAs) were once believed to lack functionality in the transcriptome due to their inability to code for proteins. LncRNAs have since been proven to be epigenetic regulators of gene expression that can establish cell phenotypes and contribute to tissue development. These RNAs can be studied to determine their functional activities in gene regulation and potentially use this knowledge to create drug targets for a variety of diseases. Several lncRNAs have been confirmed to play a role in the differentiation of mesenchymal stem cells (MSCs) to osteoblasts, which are the main bone cells necessary for creating new bone tissue. Despite the recent increase in studies relating bone formation and lncRNAs, there is still limited knowledge on specific lncRNAs proven to regulate osteogenesis. This thesis selected five lncRNAs that have been shown to be highly upregulated during osteoblast differentiation to potentially identify novel lncRNAs that play a role in bone formation. The lncRNAs were knocked down using Antisense LNA GapmeRs and the transfected MSCs were differentiated to Days 0, 3, 5, and 7 of osteoblast commitment. The relative gene expression of ALPL was determined for each knockdown lncRNA using qPCR. In addition, RNA-Seq was performed to further validate any successful findings. We determined that the knockdown of one lncRNA, LNC_7255, demonstrated reduced expression of ALPL. Gene expression profiling with RNA- Seq showed overall downregulation of genes related to ossification and extracellular matrix organization pathways. Based on the promising qPCR and RNA-Seq results, we have determined that LNC_7255 is a key regulator of osteogenesis.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Recommended Citation
Pawlowski, Michelle, "Discovery of a Novel Long Non-Coding RNA that Regulates Bone Formation" (2023). UVM Patrick Leahy Honors College Senior Theses. 583.
https://scholarworks.uvm.edu/hcoltheses/583