TY - JOUR
T1 - Inducible CRISPR genome editing platform in naive human embryonic stem cells reveals JARID2 function in self-renewal
AU - Ferreccio, Amy
AU - Mathieu, Julie
AU - Detraux, Damien
AU - Somasundaram, Logeshwaran
AU - Cavanaugh, Christopher
AU - Sopher, Bryce
AU - Fischer, Karin
AU - Bello, Thomas
AU - M. Hussein, Abdiasis
AU - Levy, Shiri
AU - Cook, Savannah
AU - Sidhu, Sonia B.
AU - Artoni, Filippo
AU - Palpant, Nathan J.
AU - Reinecke, Hans
AU - Wang, Yuliang
AU - Paddison, Patrick
AU - Murry, Charles
AU - Jayadev, Suman
AU - Ware, Carol
AU - Ruohola-Baker, Hannele
N1 - Funding Information:
We thank members of the Ruohola-Baker laboratory for helpful discussions throughout this work. We thank Kristin Goodsell, Varun Ramaswamy, Jason Miklas, and Dannie Macrin for help throughout this study. We thank Dr. Karol Bomsztyk for providing Nanog antibody. This work is supported by grants from the National Institutes of Health R01GM097372, R01GM97372-03S1 and R01GM083867 for HRB, 1P01GM081619 for CW, CM and HRB, T32CA080416 for AF, and the NHLBI Progenitor Cell Biology Consortium (U01HL099997; UO1HL099993) for PP, CW and HRB.
Funding Information:
This work was supported by the National Institute of General Medical Sciences [grant numbers P01GM081619, R01GM097372, R01GM97372-03S1 and R01GM083867]; National Cancer Institute [grant number T32CA080416]; National Heart, Lung, and Blood Institute [grant number U01HL099997].
Publisher Copyright:
© 2018 Taylor & Francis.
PY - 2018/3/4
Y1 - 2018/3/4
N2 - To easily edit the genome of naïve human embryonic stem cells (hESC), we introduced a dual cassette encoding an inducible Cas9 into the AAVS1 site of naïve hESC (iCas9). The iCas9 line retained karyotypic stability, expression of pluripotency markers, differentiation potential, and stability in 5iLA and EPS pluripotency conditions. The iCas9 line induced efficient homology–directed repair (HDR) and non-homologous end joining (NHEJ) based mutations through CRISPR-Cas9 system. We utilized the iCas9 line to study the epigenetic regulator, PRC2 in early human pluripotency. The PRC2 requirement distinguishes between early pluripotency stages, however, what regulates PRC2 activity in these stages is not understood. We show reduced H3K27me3 and pluripotency markers in JARID2 2iL-I-F hESC mutants, indicating JARID2 requirement in maintenance of hESC 2iL-I-F state. These data suggest that JARID2 regulates PRC2 in 2iL-I-F state and the lack of PRC2 function in 5iLA state may be due to lack of sufficient JARID2 protein.
AB - To easily edit the genome of naïve human embryonic stem cells (hESC), we introduced a dual cassette encoding an inducible Cas9 into the AAVS1 site of naïve hESC (iCas9). The iCas9 line retained karyotypic stability, expression of pluripotency markers, differentiation potential, and stability in 5iLA and EPS pluripotency conditions. The iCas9 line induced efficient homology–directed repair (HDR) and non-homologous end joining (NHEJ) based mutations through CRISPR-Cas9 system. We utilized the iCas9 line to study the epigenetic regulator, PRC2 in early human pluripotency. The PRC2 requirement distinguishes between early pluripotency stages, however, what regulates PRC2 activity in these stages is not understood. We show reduced H3K27me3 and pluripotency markers in JARID2 2iL-I-F hESC mutants, indicating JARID2 requirement in maintenance of hESC 2iL-I-F state. These data suggest that JARID2 regulates PRC2 in 2iL-I-F state and the lack of PRC2 function in 5iLA state may be due to lack of sufficient JARID2 protein.
KW - CRISPR-Cas9
KW - epigenetics
KW - genome editing
KW - Human embryonic stem cells
KW - JARID2
KW - naïve hESC
KW - PRC2
KW - PSEN2
UR - http://www.scopus.com/inward/record.url?scp=85045044149&partnerID=8YFLogxK
U2 - 10.1080/15384101.2018.1442621
DO - 10.1080/15384101.2018.1442621
M3 - Article
C2 - 29466914
AN - SCOPUS:85045044149
SN - 1538-4101
VL - 17
SP - 535
EP - 549
JO - Cell Cycle
JF - Cell Cycle
IS - 5
ER -