TY - JOUR
T1 - A genome-wide screen reveals new regulators of the 2-cell-like cell state
AU - Gupta, Nikhil
AU - Yakhou, Lounis
AU - Albert, Julien Richard
AU - Azogui, Anaelle
AU - Ferry, Laure
AU - Kirsh, Olivier
AU - Miura, Fumihito
AU - Battault, Sarah
AU - Yamaguchi, Kosuke
AU - Laisné, Marthe
AU - Domrane, Cécilia
AU - Bonhomme, Frédéric
AU - Sarkar, Arpita
AU - Delagrange, Marine
AU - Ducos, Bertrand
AU - Cristofari, Gael
AU - Ito, Takashi
AU - Greenberg, Maxim V.C.
AU - Defossez, Pierre Antoine
N1 - Publisher Copyright:
© 2023, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2023/8/1
Y1 - 2023/8/1
N2 - In mammals, only the zygote and blastomeres of the early embryo are totipotent. This totipotency is mirrored in vitro by mouse ‘2-cell-like cells’ (2CLCs), which appear at low frequency in cultures of embryonic stem cells (ESCs). Because totipotency is not completely understood, we carried out a genome-wide CRISPR knockout screen in mouse ESCs, searching for mutants that reactivate the expression of Dazl, a gene expressed in 2CLCs. Here we report the identification of four mutants that reactivate Dazl and a broader 2-cell-like signature: the E3 ubiquitin ligase adaptor SPOP, the Zinc-Finger transcription factor ZBTB14, MCM3AP, a component of the RNA processing complex TREX-2, and the lysine demethylase KDM5C. All four factors function upstream of DPPA2 and DUX, but not via p53. In addition, SPOP binds DPPA2, and KDM5C interacts with ncPRC1.6 and inhibits 2CLC gene expression in a catalytic-independent manner. These results extend our knowledge of totipotency, a key phase of organismal life.
AB - In mammals, only the zygote and blastomeres of the early embryo are totipotent. This totipotency is mirrored in vitro by mouse ‘2-cell-like cells’ (2CLCs), which appear at low frequency in cultures of embryonic stem cells (ESCs). Because totipotency is not completely understood, we carried out a genome-wide CRISPR knockout screen in mouse ESCs, searching for mutants that reactivate the expression of Dazl, a gene expressed in 2CLCs. Here we report the identification of four mutants that reactivate Dazl and a broader 2-cell-like signature: the E3 ubiquitin ligase adaptor SPOP, the Zinc-Finger transcription factor ZBTB14, MCM3AP, a component of the RNA processing complex TREX-2, and the lysine demethylase KDM5C. All four factors function upstream of DPPA2 and DUX, but not via p53. In addition, SPOP binds DPPA2, and KDM5C interacts with ncPRC1.6 and inhibits 2CLC gene expression in a catalytic-independent manner. These results extend our knowledge of totipotency, a key phase of organismal life.
U2 - 10.1038/s41594-023-01038-z
DO - 10.1038/s41594-023-01038-z
M3 - Article
C2 - 37488355
AN - SCOPUS:85165491231
SN - 1545-9993
VL - 30
SP - 1105
EP - 1118
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
IS - 8
ER -