Rapid RNA-protein interaction detection in living cells
Abstract/Contents
- Abstract
- RNA-protein interactions play numerous roles in cellular function and disease. Here we describe RNAprotein interaction detection (RaPID), which uses proximity-dependent protein labeling, based on the BirA* biotin ligase, to rapidly identify the proteins that bind RNA sequences of interest in living cells. RaPID displays utility in multiple applications, including in evaluating protein binding to mutant RNA motifs in human genetic disorders, in uncovering potential post-transcriptional networks in breast cancer, and in discovering essential host proteins that interact with Zika virus RNA. To improve the BirA* labeling component of RaPID, moreover, a new mutant BirA* was engineered from B. subtilis that enables > 1000-fold faster kinetics and > 30-fold increased signal-to-noise ratio over the prior standard E. coli BirA*, thereby enabling direct study of RNA-protein interactions in living cells on a timescale as short as one minute.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2017 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Muthukumar Ramanathan |
|---|---|
| Associated with | Stanford University, Interdisciplinary Stem Cell Biology and Regenerative Medicine Program. |
| Primary advisor | Khavari, Paul A |
| Thesis advisor | Khavari, Paul A |
| Thesis advisor | Oro, Anthony, 1958- |
| Thesis advisor | Sarnow, P. (Peter) |
| Advisor | Oro, Anthony, 1958- |
| Advisor | Sarnow, P. (Peter) |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Muthukumar Ramanathan. |
|---|---|
| Note | Submitted to the Interdisciplinary Stem Cell Biology and Regenerative Medicine Program. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2017. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2017 by . Muthukumar Ramanathan
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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