We generated a BPTF knockout human induced pluripotent stem cell (iPSC) line using CRISPR-Cas9 and confirmed loss of the targeted locus by genotyping. Quantitative mass spectrometry verified complete absence of BPTF protein in the knockout clone. This validated line was used for downstream functional assays in the somitoid system, a 3D iPSC-derived model that recapitulates early human somite development.