Full-length sequence of plasmids used in Y Sato and MT Hayashi, "Micronucleus is not a potent inducer of cGAS-STING pathway".

For cloning of the Sister-Control (SC) cassette plasmid (pMTH857) used for genomic integration, synthetic DNA fragments (Integrated DNA Technologies) were introduced into the original sister cassette plasmid (pMTH397)(Kagaya et al, 2020). A loxP sequence and two roxP sequences were inserted in downstream of a 5’ exon of mCitrine/mCerulean3 and neoR-franking regions, respectively, for potential future experiments. LentiCRISPR.v2 (addgene #52961) was mutagenized to introduce R691A to generate HiFi Cas9. pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (addgene #107942) was used to obtain Lenti-enAsCas12a-HF1-2C-NLS, during which one more NLS was added to the C-terminus to improve its efficiency(Liu et al, 2019). LentiGuide-puro (addgene #52963) and an improved sgRNA scaffold sequence from pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W (addgene #67979) were used to generate LentiGuide-puro-sgFUSION21-C+5bp plasmid. pH2B-miRFP703 (addgene #80001) and pCSII-EF-mVenus-hGeminin(1/110) (RDB15271) were used to generate pCSII-EF-emiRFP703-Geminin(1-110), during which the N-terminal sequence of miRFP703 was modified to obtain emiRFP703(Matlashov et al, 2020). An improved rtTA3G was artificially synthesized (Integrated DNA Technologies) to obtain pLenti-rtTA3G(Zhou et al, 2006). pCW-Cas9 (addgene #50661) was modified to generate pTRE3G-miRFP670nano-p2a-Cas9(HiFi), during which the puroR-t2a-rtTA sequence was removed. miRFP670nano was artificially synthesized (Integrated DNA Technologies)(Oliinyk et al, 2019). Mutagenesis on Cas9 and cGAS was performed by conventional PCR followed by HiFi DNA Assembly (NEB) or In-Fusion cloning (Takara Bio).