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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/31344

Title: Assessment of Cell Penetrating Peptides as a Vehicle for Delivering Transcription Factors for Stem Cell Reprogramming and Controlling Fate Decisions
Authors: Moghaddam, Bahar
Advisor: Audet, Julie
Zandstra, Peter W.
Department: Chemical Engineering and Applied Chemistry
Keywords: Cell Penetrating Peptides
Induced pluripotent stem cells
Transcription Factor Delivery
Issue Date: 14-Dec-2011
Abstract: Conjugation of the Human Immunodeficiency Virus Transactivator of Transcription (TAT) to active proteins allows transport into the intracellular environment. This feature can be harnessed to deliver combinations of reprogramming factors (RFs) such as c-Myc, Oct4, Klf4 and Sox2 into somatic cells to derive induced pluripotent stem cells (iPSCs). For this project, TAT-fusion proteins including four TAT-conjugated RFs (TAT-RFs) have been produced and purified. All four TAT-RFs can bind specific DNA sequences. Bioactivity was tested in live cells using a novel assay based on an engineered fibroblast cell line that can be induced to express RFs by doxycycline and subsequently generate iPSCs. To test each TAT-RF, reprogramming was blocked by transient silencing of a single RF by siRNA and rescued by the corresponding TAT-RF. The results of this assay suggested that TAT-Klf4 was bioactive in cells; however, definitive evidence could not be obtained for other RFs.
URI: http://hdl.handle.net/1807/31344
Appears in Collections:Master

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