Team:MIT

• 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

Splice & Dice

Using dCas13a to control alternative splicing in mammalian cells

Alternative splicing is a process that takes mRNA transcripts and modifies it in various ways to create a final mature mRNA molecule for translation. Some sequences known as introns are removed; other sequences known as exons remain to be translated. With such a process, a single gene can result in many types of proteins transcripts Our iGEM projects seeks to control alternative splicing of RNA, specifically exon skipping and inclusion, using a protein called Cas13a. This is a protein that attaches to RNA via a complementary guide RNA, then cuts the RNA strand. For our purposes, we're using a modified version of this protein, known as dCas13a, that can attach, but doesn't cut. By targeting certain portions of a fluorescent protein construct, we can determine whether or not we achieved the intended isoforms based on the presence or absence of the fluorescent protein in addition to sequencing.