Team:MIT/REST

• ABOUT US
  Team Attributions
• BACKGROUND
  Alternative Splicing CRISPR RNA Binding Proteins REST
• PROJECT
  Guides and ASOs mKate-FF4 2 Exon mKate HBG 3 Exon mKate HBG 3 Exon Dual Fluorescence
• LAB WORK
  Parts/Improved Parts Protocols Notebook Future Work
• MODELLING
• HUMAN PRACTICES
  Integrated HP Public Engagement Collaborations InterLab
• AWARDS
  HP Silver HP Gold Education & Public Engagement Model Parts Collection

We have selected misregulation of the transcription factor REST in certain forms of aggressive cancer as a disease state that could potentially be treated with our system for alternative splicing regulation. REST protein, encoded by the REST gene, is a tumor suppressor protein that binds to more than 2000 known target oncogenes. (1) It represses transcription by binding a DNA sequence element called the neuron-restrictive silencer element, known as RE1, recruiting co-repressors as well as chromatin modifying enzymes that change the topology of the oncogenes, all in all down-regulating the expression of these oncogenes.

In a normal splicing event, exon 4 in the pre-mature mRNA is skipped and the mature mRNA is accordingly translated. A certain balance of isoforms from the REST gene is required for functional suppression function.

< a href="https://static.igem.org/mediawiki/parts/c/cd/Fxnal_REST.png">

Loss of REST protein has been linked to forms of SCLC, colon cancer, and recently researchers at Wisconsin University have discovered a subset of breast cancers which display a highly aggressive disease course in which the tumor suppressor gene REST is lost. (2) In this subset, REST gene is mis-spliced, leading to the inclusion of the normally skipped exon. This exon includes a premature stop codon, which, upon its identification by the translation machinery, results in the production of a truncated REST protein called REST4.

The new protein also lacks a C terminus for recruiting the corepressors and chromatin modifying enzymes, leading to the protein’s loss of function as a tumor suppressor.