Here, the scientists have developed a high throughput CRISPR/Cas9-based genome-editing strategy and used it to interrogate 174 candidate regulatory sequences within the 1Mbp POU5F1 locus in the human embryonic stem cells (hESCs). They identified two classical regulatory elements that are essential for POU5F1 transcription in the hESCs. Unexpectedly, they also discovered a new class of enhancers that contribute to POU5F1 transcription in an unusual way: disruption of such sequences led to a temporary loss of POU5F1 transcription that is fully restored after a few rounds of cell division. These results demonstrate the utility of a high throughput screening for functional characterization of non-coding DNA, and reveal a previously unrecognized layer of gene regulation in human cells.
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Here, the scientists have developed a high throughput CRISPR/Cas9-based genome-editing strategy and used it to interrogate 174 candidate regulatory sequences within the 1Mbp POU5F1 locus in the human embryonic stem cells (hESCs). They identified two classical regulatory elements that are essential for POU5F1 transcription in the hESCs. Unexpectedly, they also discovered a new class of enhancers that contribute to POU5F1 transcription in an unusual way: disruption of such sequences led to a temporary loss of POU5F1 transcription that is fully restored after a few rounds of cell division. These results demonstrate the utility of a high throughput screening for functional characterization of non-coding DNA, and reveal a previously unrecognized layer of gene regulation in human cells.