Rebecca Lea Poster BSDB

The transcriptional regulatory circuitry underlying human pluripotency is still unclear. Bona fide pluripotency exists in the preimplantation epiblast, which gives rise to embryonic tissues and hESCs. However, conventional hESCs represent a state of “primed” pluripotency. “Naïve” hESCs, more transcriptionally and epigenetically akin to human epiblast, have been developed. Recent single-cell transcriptome analyses of human embryos have enabled us to identify novel transcription factors, including the zinc-finger protein KLF17, that are enriched in the human epiblast and activated in naïve hESCs. We are investigating a putative role for KLF17 in regulating human pluripotency.

Using ectopic expression of KLF17 in conventional hESCs, we have shown KLF17 to be sufficient to upregulate other naïve-specific genes. That KLF17 alone activates a more naïve profile in primed culture conditions is suggestive of a functional role.

However, using CRISPR/Cas9-mediated knockout of KLF17 prior to resetting to naïve pluripotency, we have observed that KLF17 is not required for naïve pluripotency in vitro, as KLF17-knockout cells survive for at least 10 passages. We are investigating the transcriptional profile of wild-type and KLF17-KO naïve hESCs to identify any genes that are misexpressed upon KLF17 loss-of-function.

Through these studies, we hope to elucidate any role of KLF17 in naïve pluripotency in vitro.