Enhancing mRNA stability through the addition of stabilizing untranslated regions

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.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Mechanical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2011

Author dc:creator

• Tran, Thi (Thi Thi Kim)

Advisor dc:contributor.advisor

• Mehmet F. Yanik.

• Mechanical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/68694

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/68694