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| Content Provider | IEEE Xplore Digital Library |
|---|---|
| Author | Overby, D.R. Alenghat, F.J. Montoya-Zavala, M. HuCheng Bei Philmo Oh Karavitis, J. Ingber, D.E. |
| Copyright Year | 1965 |
| Abstract | This paper focuses on the development of magnetic cellular switches to enable magnetic control of intracellular functions in living mammalian cells, including receptor signal transduction and gene transcription. Our approach takes advantage of the mechanosensitivity of adenosine 3',5'-monophosphate (cAMP) induction and downstream transcription controlled by the cAMP regulatory element (CRE) to engineer gene constructs that optically report gene expression in living cells. We activate transcription of these gene reporters by applying magnetic (mechanical) stress to magnetic microbeads bound to cell surface integrin receptors. In these gene reporter constructs, CRE motifs drive the expression of fluorescent proteins or enzymes that produce fluorescent products, such as DsRed and /spl beta/-lactamase (BLA), respectively. We demonstrate that a chemical inducer of cAMP (forskolin) increases expression of CRE-DsRed in living cells. More importantly, a threefold increase in CRE-BLA expression is induced by application of mechanical stress to magnetic microbeads (4.5 /spl mu/m) bound to cell surface integrin receptors. Induction of cAMP could be detected within 5 min using a protein fragment complementation assay involving interactions between the KID and KIX domains of the CRE binding protein linked to complementary halves of the BLA enzyme. These studies confirm that application of magnetic stress to integrins induces gene transcription by activating the cAMP-dependent transcription factor CREB. Ongoing studies focus on optimizing sensitivity and reducing signal-to-noise by establishing stable cell lines that express these gene reporters. These studies collectively demonstrate the feasibility of using magnetic technologies to control function in living mammalian cells and, hence, support the possibility of developing magnetically-actuated cellular components for use in future micro- and nanotechnologies. |
| Sponsorship | IEEE Magnetics Society |
| Starting Page | 2958 |
| Ending Page | 2960 |
| Page Count | 3 |
| File Size | 125244 |
| File Format | |
| ISSN | 00189464 |
| Volume Number | 40 |
| Issue Number | 4 |
| Language | English |
| Publisher | Institute of Electrical and Electronics Engineers, Inc. (IEEE) |
| Publisher Date | 2004-07-01 |
| Publisher Place | U.S.A. |
| Access Restriction | One Nation One Subscription (ONOS) |
| Rights Holder | Institute of Electrical and Electronics Engineers, Inc. (IEEE) |
| Subject Keyword | Magnetic switching Stress Micromagnetics Fluorescence Biochemistry Proteins Optical switches Optical control Optical sensors Gene expression |
| Content Type | Text |
| Resource Type | Article |
| Subject | Electronic, Optical and Magnetic Materials Electrical and Electronic Engineering |
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