Figure 1.
Engineering of a STING1-V155M K562 model and functional assessment of bespoke base and prime editing. (A) Bidirectional third-generation lentiviral (LV) vector used to generate stable K562wt and K562mut lines expressing STING1-WT or STING1-V155M together with an EGFRt surface marker. (B) In vitro transcription (IVT) mRNA electroporation of top guide–editor pairs yields high A-to-G editing in K562mut, including base editing with sgRNA2/sgRNA5, a bystander-reducing TadA variant (var17) with sgRNA5, and prime editing with PEGsm in PE2 and PE3 configurations; editing was quantified by Sanger sequencing (EditR). (C) Functional readout by ddPCR showing IFIT1 and ISG15 expression (normalized to HPRT1) in K562mut across editing conditions and after STING inhibition (H-151). Statistics were computed by one-way ANOVA with multiple comparisons correction; significance is indicated, and where not otherwise specified, **** denotes p < 0.0001. Refer to the image caption for details.

Engineering of a STING1-V155M K562 model and functional assessment of bespoke base and prime editing. (A) Bidirectional third-generation lentiviral (LV) vector used to generate stable K562wt and K562mut lines expressing STING1-WT or STING1-V155M together with an EGFRt surface marker. (B) In vitro transcription (IVT) mRNA electroporation of top guide–editor pairs yields high A-to-G editing in K562mut, including base editing with sgRNA2/sgRNA5, a bystander-reducing TadA variant (var17) with sgRNA5, and prime editing with PEGsm in PE2 and PE3 configurations; editing was quantified by Sanger sequencing (EditR). (C) Functional readout by ddPCR showing IFIT1 and ISG15 expression (normalized to HPRT1) in K562mut across editing conditions and after STING inhibition (H-151). Statistics were computed by one-way ANOVA with multiple comparisons correction; significance is indicated, and where not otherwise specified, **** denotes p < 0.0001.

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