Project
Project #2391
Step by step one goes very far
Organisms :
Group : Name of Applicant : Pablo Navarro Gil Date of application : 09-02-2015 Unit : Epigenetics of Stem Cells Location : Monod (66) – 2eme etage Phone : 0145688285@ Mail : pablo.navarro-gil@pasteur.fr
Project context and summary :
We have recently identified around 500 long non-coding RNAs that follow a very precise expression pattern in undifferentiated mouse Embryonic Stem cells. In order to assess if any of those molecules are functionally relevant for the biology of ES cells we will perform a functional screening based on the CRISPR technology. We plan to use a modified version of the CRISPR-CAS9 technology, the CRISPRon system, in which (i) the guide-RNAs (gRNAs) will target the promoters of our candidate lncRNAs (at least 5 gRNAs per promoter) and (ii) an enzymatically-inactive CAS9 will be engineered to recrute 10 molecules of the potent VP64 transactivator. This will enable to upregulate the targeted lncRNA from its endogeneous locus.
Related team publications :