Editing repeats in Huntington's:fewer somatic repeat expansions in patient cells

This research complies with relevant ethical regulations. The study protocol was approved by the Broad’s Institutional Biosafety Committee, the Broad’s Institutional Animal Care and Use Committee (IACUC) and relevant IACUC compliance committees at Massachusetts General Hospital. Cell culture Culture of mESCs and HEK293T cells was performed according to previously published protocols100. Undifferentiated 129P2/OlaHsd mESC (male) lines125 were maintained on 0.2% gelatin-coated plates feeder-free in mESC media composed of Knockout DMEM (Life Technologies) supplemented with 15% defined FBS (HyClone), 0.1 mM nonessential amino acids (Life Technologies), Glutamax (Life Technologies), 0.55 mM 2-mercaptoethanol (Sigma-Aldrich), 1X ESGRO LIF (Millipore), with the addition of 2i: 5 nM GSK-3 inhibitor XV (Sigma-Aldrich), and 500 nM UO126 (Sigma-Aldrich). FXN-mESCs were generated for this project (see below). HEK293T cells were purchased from ATCC (CRL-3216) and were maintained in DMEM (Thermo F ... Read full article.