Recombinase-based conditional and reversible gene regulation via XTR alleles
Synthetic biological tools that enable precise regulation of gene function within in vivo systems have enormous potential to discern gene function in diverse physiological settings. Here we report the development and characterization of a synthetic gene switch that, when targeted in the mouse germli...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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2016
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| Online Access: | http://hdl.handle.net/1721.1/100823 https://orcid.org/0000-0001-5785-8911 |
| _version_ | 1826214270371627008 |
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| author | Robles-Oteiza, Camila Yates, Travis Cicchini, Michelle Lauderback, Brian Burds, Aurora A. Winslow, Monte M. Feldser, David M. Taylor, Sarah E. Cashman, Chris Jacks, Tyler E |
| author2 | Koch Institute for Integrative Cancer Research at MIT |
| author_facet | Koch Institute for Integrative Cancer Research at MIT Robles-Oteiza, Camila Yates, Travis Cicchini, Michelle Lauderback, Brian Burds, Aurora A. Winslow, Monte M. Feldser, David M. Taylor, Sarah E. Cashman, Chris Jacks, Tyler E |
| author_sort | Robles-Oteiza, Camila |
| collection | MIT |
| description | Synthetic biological tools that enable precise regulation of gene function within in vivo systems have enormous potential to discern gene function in diverse physiological settings. Here we report the development and characterization of a synthetic gene switch that, when targeted in the mouse germline, enables conditional inactivation, reports gene expression and allows inducible restoration of the targeted gene. Gene inactivation and reporter expression is achieved through Cre-mediated stable inversion of an integrated gene-trap reporter, whereas inducible gene restoration is afforded by Flp-dependent deletion of the inverted gene trap. We validate our approach by targeting the p53 and Rb genes and establishing cell line and in vivo cancer model systems, to study the impact of p53 or Rb inactivation and restoration. We term this allele system XTR, to denote each of the allelic states and the associated expression patterns of the targeted gene: eXpressed (XTR), Trapped (TR) and Restored (R). |
| first_indexed | 2024-09-23T08:14:24Z |
| format | Article |
| id | mit-1721.1/100823 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:14:24Z |
| publishDate | 2016 |
| record_format | dspace |
| spelling | mit-1721.1/1008232022-09-30T08:32:26Z Recombinase-based conditional and reversible gene regulation via XTR alleles Robles-Oteiza, Camila Yates, Travis Cicchini, Michelle Lauderback, Brian Burds, Aurora A. Winslow, Monte M. Feldser, David M. Taylor, Sarah E. Cashman, Chris Jacks, Tyler E Koch Institute for Integrative Cancer Research at MIT Taylor, Sarah Cashman, Christopher R. Burds, Aurora A. Jacks, Tyler E. Synthetic biological tools that enable precise regulation of gene function within in vivo systems have enormous potential to discern gene function in diverse physiological settings. Here we report the development and characterization of a synthetic gene switch that, when targeted in the mouse germline, enables conditional inactivation, reports gene expression and allows inducible restoration of the targeted gene. Gene inactivation and reporter expression is achieved through Cre-mediated stable inversion of an integrated gene-trap reporter, whereas inducible gene restoration is afforded by Flp-dependent deletion of the inverted gene trap. We validate our approach by targeting the p53 and Rb genes and establishing cell line and in vivo cancer model systems, to study the impact of p53 or Rb inactivation and restoration. We term this allele system XTR, to denote each of the allelic states and the associated expression patterns of the targeted gene: eXpressed (XTR), Trapped (TR) and Restored (R). 2016-01-14T00:28:20Z 2016-01-14T00:28:20Z 2015-11 Article http://purl.org/eprint/type/JournalArticle 2041-1723 http://hdl.handle.net/1721.1/100823 Robles-Oteiza, Camila, Sarah Taylor, Travis Yates, Michelle Cicchini, Brian Lauderback, Christopher R. Cashman, Aurora A. Burds, Monte M. Winslow, Tyler Jacks, and David M. Feldser. “Recombinase-Based Conditional and Reversible Gene Regulation via XTR Alleles.” Nat Comms 6 (November 5, 2015): 8783. © 2015 Macmillan Publishers Limited https://orcid.org/0000-0001-5785-8911 en_US http://dx.doi.org/10.1038/ncomms9783 Nature Communications Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Nature Publishing Group |
| spellingShingle | Robles-Oteiza, Camila Yates, Travis Cicchini, Michelle Lauderback, Brian Burds, Aurora A. Winslow, Monte M. Feldser, David M. Taylor, Sarah E. Cashman, Chris Jacks, Tyler E Recombinase-based conditional and reversible gene regulation via XTR alleles |
| title | Recombinase-based conditional and reversible gene regulation via XTR alleles |
| title_full | Recombinase-based conditional and reversible gene regulation via XTR alleles |
| title_fullStr | Recombinase-based conditional and reversible gene regulation via XTR alleles |
| title_full_unstemmed | Recombinase-based conditional and reversible gene regulation via XTR alleles |
| title_short | Recombinase-based conditional and reversible gene regulation via XTR alleles |
| title_sort | recombinase based conditional and reversible gene regulation via xtr alleles |
| url | http://hdl.handle.net/1721.1/100823 https://orcid.org/0000-0001-5785-8911 |
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