2026-09-19T09:05:57Zhttps://dspace.mit.edu/server/oai/requestoai:dspace.mit.edu:1721.1/686942022-01-13T07:54:36Zcom_1721.1_7582com_1721.1_7581col_1721.1_131024
Mehmet F. Yanik.
Tran, Thi (Thi Thi Kim)
Massachusetts Institute of Technology. Dept. of Mechanical Engineering.
Massachusetts Institute of Technology. Department of Mechanical Engineering
2012-01-30T15:21:50Z
2012-01-30T15:21:50Z
2011
2011
http://hdl.handle.net/1721.1/68694
773197160
Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2011.
This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.
Cataloged from student submitted PDF version of thesis.
Includes bibliographical references (p. 20).
Messenger-RNA (mRNA) therapy, in which mRNA is introduced to cells or tissues to cause a transient expression of specific genes, has applications ranging from tissue engineering to neural regeneration. This transient nature of mRNA, however, limits the duration of gene upregulation, and potentially reduces the efficacy of mRNA therapy. In this paper, various mRNA constructs were studied to identify stable designs which could be used for long-lasting mRNA therapies. Specifically, the GFP gene was prepared with four different hemoglobin untranslated regions (UTR's), which are known for their stabilization effects on mRNA. Each mRNA template was created through the digestion and ligation of a cDNA template, reversed transcribed from total RNA, and transfected into fibroblasts. The resulting fluorescence was measured as a surrogate for translation efficiency and duration. It was determined that the human beta-globin B UTR resulted in a fluorescence level that was tenfolds brighter than human alpha-globin 1 (HBA1), which was the least effective stabilizing untranslated region. This highlights the importance of UTR selection for future mRNA therapies.
by Thi Tran.
S.B.
20 p.
eng
Massachusetts Institute of Technology
M.I.T. theses are protected by
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http://dspace.mit.edu/handle/1721.1/7582
Mechanical Engineering.
Enhancing mRNA stability through the addition of stabilizing untranslated regions
Thesis
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<Abstract>Messenger-RNA (mRNA) therapy, in which mRNA is introduced to cells or tissues to cause a transient expression of specific genes, has applications ranging from tissue engineering to neural regeneration. This transient nature of mRNA, however, limits the duration of gene upregulation, and potentially reduces the efficacy of mRNA therapy. In this paper, various mRNA constructs were studied to identify stable designs which could be used for long-lasting mRNA therapies. Specifically, the GFP gene was prepared with four different hemoglobin untranslated regions (UTR's), which are known for their stabilization effects on mRNA. Each mRNA template was created through the digestion and ligation of a cDNA template, reversed transcribed from total RNA, and transfected into fibroblasts. The resulting fluorescence was measured as a surrogate for translation efficiency and duration. It was determined that the human beta-globin B UTR resulted in a fluorescence level that was tenfolds brighter than human alpha-globin 1 (HBA1), which was the least effective stabilizing untranslated region. This highlights the importance of UTR selection for future mRNA therapies.</Abstract>
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