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The fate of the X chromosome in cells differentiated from female human embryonic stem 
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BigField GEG Tech's insight:
The authors inactivated all three catalytically active DNA methyltransferases (DNMTs) in human embryonic stem cells (ESCs) using CRISPR/Cas9 genome editing. Surprisingly, in contrast to findings in mouse, deletion of DNMT1 resulted in rapid cell death in human ESCs. To overcome this immediate lethality, they generated a doxycycline-responsive tTA-DNMT1 rescue line and readily obtained homozygous DNMT1-mutant lines. However, doxycycline-mediated repression of exogenous DNMT1 initiates rapid, global loss of DNA methylation, followed by extensive cell death.
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Scoop.it!
BigField GEG Tech's insight:
This study demonstrates a knockin strategy without drug selection for both active and silent genes in human embryonic stem cells (hESCs) thanks to the iCRISPR platform. This selection-free knockin strategy is expected to greatly facilitate the use of hESCs for developmental studies, disease modeling, and cell-replacement therapy.
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Here, the scientists demonstrate that X chromosome dosage compensation is not required for ESC differentiation. Their data imply that XiXIST+Xa ESCs are most suited for downstream applications and show that all other X states are abnormal byproducts of their ESC derivation and propagation method.