Generation and validation of versatile inducible CRISPRi embryonic stem cell and mouse model
Fig 2
Induced dCas9-KRAB at active gene promoters by sgRNAs or multi-gRNAs.
(A, B) RT-qPCR analysis of stable iKRAB ESCs containing sgRNA against Oct4 or multi-gRNAs against Oct4 or Nanog after 2 days of Dox induction. The binding location of each gRNA is indicated relative to the TSS of Oct4 or Nanog locus. (C) The cell morphology of Oct4 or Nanog multi-gRNAs-transduced iKRAB ESCs after 2 days of Dox induction. The scale bar represents 100 μm. The numerical values used to generate graphs in panels A and B are available in S1 Data. Cas, CRISPR-associated; dCas9, deactivated Cas9; Dox, doxycycline; ESC, embryonic stem cell; KRAB, Krüppel-associated box; RT-qPCR, reverse transcription PCR; sgRNA, single-guide RNA; TSS, transcription start site.