Periodic Reporting for period 1 - ScrIntOR (Genome-wide CRISPR/Cas9 screen in intestinal organoids for identification and molecular characterization of therapeutic targets to enhance tissue regeneration)
Okres sprawozdawczy: 2021-05-01 do 2023-04-30
In this project, we will combine in vitro intestinal organoid cultures with a CRISPR/Cas9 screening approach to (i) identify novel regulators of regeneration in the intestinal epithelium and (ii) decipher how this process is controlled at the molecular level. The outcome of the screen and the downstream validation will substantially enhance the current understanding of intestinal repair upon tissue damage and is likely to aid in the identification of molecular pathways that will constitute new therapeutic targets for patients suffering from IBD.
Within the reporting period, I have been successfully working on the characterization of intestinal organoid cultures as well as establishing the set-up for the genome-wide CRISPR/Cas9 screen, and I will be soon ready to perform the actual screen. I have therefore established intestinal organoid cultures under conventional conditions as well as grown in a bioreactor system, which will allow simultaneous growth of millions of organoids. These cultures upon appropriate stimulation can enter a regenerative state. In addition, I have generated intestinal epithelial cell lines which express Cas9 in an inducible manner.
Due to early termination of the MSCA fellowship, no exploitation or dissemination of the results was planned at this stage yet.
Nevertheless, the project combines state-of-the-art organoid cultures with a genome-wide CRISPR/Cas9 screening approach intending to identify the molecular drivers of intestinal regeneration. The results of the screen will be already a significant resource on their own, however, we are aiming to move beyond a purely descriptive approach by dissecting the molecular events that orchestrate tissue repair and validating critical pathway components in a colitis mouse model. Thus, this study will potentially facilitate the identification of novel drug targets for patients suffering from IBD.