Advanced Search
DSpace@MIT

Extended transient transfection by repeated delivery of in vitro-transcribed RNA

Research and Teaching Output of the MIT Community

Show simple item record

dc.contributor.advisor Mehmet F. Yanik. en_US
dc.contributor.author Angel, Matthew en_US
dc.contributor.other Massachusetts Institute of Technology. Dept. of Electrical Engineering and Computer Science. en_US
dc.date.accessioned 2009-10-01T16:00:02Z
dc.date.available 2009-10-01T16:00:02Z
dc.date.copyright 2008 en_US
dc.date.issued 2008 en_US
dc.identifier.uri http://hdl.handle.net/1721.1/47895
dc.description Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2008. en_US
dc.description Includes bibliographical references (p. 53-56). en_US
dc.description.abstract An experimental study was performed to evaluate a novel method of controlling protein expression by repeatedly transfecting cells with in vitro-transcribed RNA. Transcripts encoding six factors, each known to play an important role in cell-type specification and maintenance, were designed and synthesized. Aspects of the design were optimized, and the intracellular stability and translation efficiency of the ivT RNA were quantified. Transcripts were delivered to cells by electroporation, and a method of increasing the rate at which cells recover from ivT-RNA transfection by combined knockdown of innate-immune-related genes was developed. Using this technique a high, approximately steady-state level of protein expression was transduced in MRC-5 human fetal-lung fibroblasts by repeated transfection with capped, poly(A)+ ivT RNA encoding a protein with an intracellular half-life of approximately three days. Transfection at 24-hour intervals with ivT RNA encoding a less stable protein resulted in protein expression that peaked twelve hours after each transfection, and diminished before the next transfection. In both cases, cells sustained a high rate of proliferation. In this study, extended transient transfection by repeated delivery of ivT RNA was shown to transduce expression of defined factors in cultured cells without genetic modification or the extensive screening required in small-molecule approaches and with significant control over the level of transduced protein. This technique may become a powerful tool in the development of new directed-differentiation and cell-type-conversion protocols. en_US
dc.description.statementofresponsibility by Matthew Angel. en_US
dc.format.extent 56 p. en_US
dc.language.iso eng en_US
dc.publisher Massachusetts Institute of Technology en_US
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. en_US
dc.rights.uri http://dspace.mit.edu/handle/1721.1/7582 en_US
dc.subject Electrical Engineering and Computer Science. en_US
dc.title Extended transient transfection by repeated delivery of in vitro-transcribed RNA en_US
dc.type Thesis en_US
dc.description.degree S.M. en_US
dc.contributor.department Massachusetts Institute of Technology. Dept. of Electrical Engineering and Computer Science. en_US
dc.identifier.oclc 435528760 en_US


Files in this item

Name Size Format Description
435528760.pdf 24.71Mb PDF Preview, non-printable (open to all)
435528760-MIT.pdf 24.71Mb PDF Full printable version (MIT only)

This item appears in the following Collection(s)

Show simple item record

MIT-Mirage