Research Article

Generation of Inducible CRISPRi and CRISPRa Human Stromal/Stem Cell Lines for Controlled Target Gene Transcription during Lineage Differentiation

Figure 3

Dynamic regulation of ALP expression during osteoblast differentiation in inducible CRISPRi or CRISPRa hMSC. Inducible CRISPRi (a) or CRISPRa (b) hMSC were transfected with ALP gRNA oligo by DharmaFECT™ Transfection Reagent and cultured with increasing concentrations of Dox. ALP expression was measured by RT-qPCR at day 2. Inducible CRISPRi (c) or CRISPRa (d) hMSC was transfected with ALP gRNA oligo or negative control (Ctrl) and induced to OB differentiation in the absence or presence of Dox (1000 ng/ml). ALP activity and staining (bottom photomicrographs) were performed on day 7 in hMSC-CRISPRi or day 3 in hMSC-CRISPRa to show the most evident change for inhibition or activation. Data are expressed as the . , , and .
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