RETROFIT_101018670

Title RETROFIT_101018670
Acronym HERVH-HESC
Start date 2021-06-01
End date
Sponsor European Research Council (ERC)
Institution The Francis Crick Institute Limited

Associated cell lines

Project Description

HESC pluripotency is a complex, highly regulated phenotype that involves expression of a number of critical transcription factors and their target genes, not all of which are currently known. Whist gene expression patterns and their correlation with pluripotency can be derived from existing expression profiling (e.g. RNA-seq data) from which function can be inferred, validating the role of any factor in HESC pluripotency will ultimately require a genetic experiment. We have identified a regulatory network driven by human endogenous retrovirus H (HERVH) that, we believe, participates in the maintenance of HESC pluripotency. We aim to manipulated this network firstly by generating genetic deficiency in a specific HERVH-initiated transcript (using Cas9-mediated gene editing) and secondly by overexpressing this transcript in HESC lines. The readout will be changes in the HESC pluripotency (assessed by RNA and protein expression of key pluripotency markers in genetically altered HESC lines. Preliminary data with immortalised somatic cell lines have provided support for the role of the identified HERVH-initiated transcript in stemness and differentiation and validated the technical aspects of Cas9-medeated editing and expression vectors. Together, these experiments should establish whether the identified HERVH-initiated transcript is necessary and sufficient to drive HESC pluripotency.