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. Author manuscript; available in PMC: 2023 May 1.
Published in final edited form as: Nat Biomed Eng. 2022 Aug 8;6(11):1284–1297. doi: 10.1038/s41551-022-00915-0

Extended Data Fig. 2. Generation, validation, and differentiation of TRAC−/−-TiPS.

Extended Data Fig. 2.

a, CRISPR/Cas9-targeted integration of EF1a-GFP-P2A-Puromycing-bGHpA (G2AP) expression unit into the TRAC locus. Top, TRAC locus; middle, plasmid containing the G2AP expression unit flanked by homology arms; bottom, edited TRAC locus. ‘FWD’ and ‘REV’ indicate the location of the forward and reverse primers used in b. b, PCR validation of G2AP integration into the TRAC locus of TiPS clones. c, Flow cytometric analysis of pluripotency marker expression on TRAC−/−-TiPS. Gated on live cells. d, T lymphoid makers of WT-TiPS and TRAC−/−-TiPS during differentiation on OP9-mDLL1 at the indicated timepoints. Gated on live CD45+ cells. D40 is as presented in Fig. 1c.