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. Author manuscript; available in PMC: 2026 Apr 2.
Published in final edited form as: Acta Biomater. 2024 Nov 8;191:276–291. doi: 10.1016/j.actbio.2024.11.007

Fig. 2. Targeted Regulation of ZNF865 Increases Cell Proliferation in ACAN/Col2a1 hASCs.

Fig. 2.

(A) The dCas9-VPR CRISPRa system acts like a synthetic transcription factor increasing expression of target genes. (B) Initial guide screening verifies ZNF865 upregulation and displays our ability to titrate target gene regulation (n = 4, *=p < 0.05). (C) Targeted upregulation of ZNF865 results in a strong correlation of increased proliferation rates in hASCs, (D) which corresponds with a shift in cell cycle in ZNF865-engienered hASCs compared to NTC-hASCs. (E) qRT-PCR was used to monitor changes in ZNF865 expression over 8-weeks, showing increasing expression to week 4 and decreasing expression back to baseline levels by week 8 (n = 3–4, *=p < 0.05). (F) qRT-PCR verifies the upregulation of (D) Col2a1 and (G) ACAN in ACAN/Col2a1-NTC and ACAN/Col2a1-ZNF865-edited hASCs compared to naïve hASC baseline expression (n = 4, *=comparing to VPR-NTC, #=comparing to ACAN/Col2a1-NTC, *,#=p < 0.05). (H) The resulting targeted regulation of ZNF865 in ACAN/Col2a1 hASCs increases proliferation rates resulting in decreased doubling time (n = 4, *=p < 0.05). NTC is nontarget control.