Genetic_screening__of_human_stem_cell_differentiation_for_endodermal_cell_lineage-sc-3458
Genetic_screening__of_human_stem_cell_differentiation_for_endodermal_cell_lineage
This study will analyse the guide sequence which were used for making mutations in the Cas9-expressing cells. We used GeCKO v2 library which were released by Feng Zhang, 2014.  This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
The goal of this project is to investigate genes regulating definitive endodermal differentiation of human iPSC using genetic screening. The development of definitive endoderm (DE) and its subsequent patterning and differentiation leads to the formation of many of the major organs including the liver, pancreas, lungs, thyroid and intestines. Efficient differentiation to DE is essential for therapeutic purpose, such as transplantation. In recent studies, specific growth factors are used to generate definitive endodermal cells from ESCs including nodal signaling pathway. Although DE differentiation methods using growth factors are useful strategies for generating DE with the ability to differentiate into hepatic or pancreatic lineages, they are not efficient enough for generation of homogenous DE populations. To improve the DE differentiation efficacy, there has been several attempt to find critical genes including transcription factors for DE differentiation. We strongly believe that a more detailed understanding of the molecular pathway of human iPS cell differentiation into DE will help the optimization of differentiation protocol. For this goal, we intend to develop an efficient phenotype-based screening strategy using genome-wide CRISPR gRNA library and deep sequencing to identify genes in DE differentiation. Furthermore, this phenotype-based screening approach can be extended into investigation for further differentiation process like a liver and pancreas progenitors from definitive endodermal cells.