Figure 1.
CRISPRi viability screens identify MIR17HG as a leading cell growth dependency in MM. (A) Schematic of CRISPRi viability screens. (B) Robust rank algorithm (RRA)-based ranked analysis of lncRNA dependencies in the secondary screen, considering 4 MM cell lines either together or individually. The top lncRNA dependency, MIR17HG, is highlighted, along with the protein-coding genes IRF4 and MYC used as positive controls. (C) CCK-8 proliferation assay of MM cell lines (AMO1, H929, KMS11, and KMS12BM) stably expressing KRAB-dCAS9 fusion protein and transduced with lentivectors to conditionally express anti-MIR17HG sgRNAs. CCK-8 assay was performed at indicated time points after exposure to doxycycline (0.5 μg/mL). Cell proliferation is calculated compared with parental cells infected with the empty sgRNA vector and exposed to doxycycline under the same conditions. (D) CCK-8 proliferation assay of MM cell lines (n = 11) transfected with 2 different ASOs targeting the MIR17HG pre-RNA or a nontargeting ASO (NC). ASOs were used at a concentration of 25 nM. Cell viability was measured 2 and 4 days after electroporation, and it is represented as % viability compared with cells transfected with NC-ASO. Data from 1 out of 3 independent experiments are shown in panel D. Data present mean ± standard deviation in panel D. ∗P < .05 by Student t test.

CRISPRi viability screens identify MIR17HG as a leading cell growth dependency in MM. (A) Schematic of CRISPRi viability screens. (B) Robust rank algorithm (RRA)-based ranked analysis of lncRNA dependencies in the secondary screen, considering 4 MM cell lines either together or individually. The top lncRNA dependency, MIR17HG, is highlighted, along with the protein-coding genes IRF4 and MYC used as positive controls. (C) CCK-8 proliferation assay of MM cell lines (AMO1, H929, KMS11, and KMS12BM) stably expressing KRAB-dCAS9 fusion protein and transduced with lentivectors to conditionally express anti-MIR17HG sgRNAs. CCK-8 assay was performed at indicated time points after exposure to doxycycline (0.5 μg/mL). Cell proliferation is calculated compared with parental cells infected with the empty sgRNA vector and exposed to doxycycline under the same conditions. (D) CCK-8 proliferation assay of MM cell lines (n = 11) transfected with 2 different ASOs targeting the MIR17HG pre-RNA or a nontargeting ASO (NC). ASOs were used at a concentration of 25 nM. Cell viability was measured 2 and 4 days after electroporation, and it is represented as % viability compared with cells transfected with NC-ASO. Data from 1 out of 3 independent experiments are shown in panel D. Data present mean ± standard deviation in panel D. ∗P < .05 by Student t test.

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