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. Author manuscript; available in PMC: 2022 Apr 14.
Published in final edited form as: Oncogene. 2021 Oct 14;40(47):6527–6539. doi: 10.1038/s41388-021-02053-4

Figure 3: MYCASOs downregulate MYC expression through RNase H1-mediated cleavage of MYC mRNA.

Figure 3:

(a) MYCASOs induce cleavage of MYC mRNA. Top: schematic of 5’ Rapid Amplification of cDNA Ends (RACE) assay. Gene specific primers (GSPs) are used to capture and amplify MYC mRNA fragments from total cellular RNA for visualization on an agarose gel. Bottom: 5’ RACE PCR products following MYCASO treatment. MYCASO-treated samples contain fragments consistent with mRNA cleavage at MYCASO binding sites, whereas vehicle- and NT-ASO-treated cells produce no fragments. HeLa cells were treated with 10 nM MYCASO in Lipofectamine for 24 hours. CUTLL1, MM1.S, and HLF cells were treated with 10 μM MYCASO (gymnotic) for 72 hours (CUTLL1 and MM1.S) or 120 hours (HLF). (b) RNase H1 knockdown rescues MYCASO-mediated MYC mRNA knockdown. Top: RT-PCR for RNASEH1 in HeLa cells following treatment with an anti-RNASEH1 ASO (ISIS194178) in Lipofectamine for 24 hours normalized to vehicle-treated control. Bottom: RT-PCR for MYC in HeLa cells following co-transfection with the indicated doses of anti-RNASEH1 ASO and MYCASO-3 in Lipofectamine 2000 for 24 hours normalized to vehicle-treated control. Values represent quadruplicate means +/− SD.