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Nickase fidelity drives EvolvR-mediated diversification in mammalian cells.

Hurtado JE, Schieferecke AJ, Halperin SO, Guan J, Aidlen D, Schaffer DV, Dueber JE
Nat Commun. 2025 Apr 19;16(1):3723. doi: 10.1038/s41467-025-58414-0. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
249060pEvolvR-mCherry-enCas9(D10A)-PolI5MExpresses nucleus-localized enCas9 (D10A) fused to PolI5M and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249061pEvolvR-mCherry-enCas9(D10A)-PolI3MExpresses nucleus-localized enCas9 (D10A) fused to PolI3M and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249063p-mCherry-enCas9(D10A)Expresses nucleus-localized enCas9 (D10A) fused to mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249064pEvolvR-mCherry-enCas9(H840A)-PolI5MExpresses nucleus-localized enCas9 (H840A) fused to PolI5M and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249065p-mCherry-dCas9-PolI5MExpresses nucleus-localized dCas9 fused to PolI5M and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249066pEvolvR-mCherry-nCas9-PolI5MExpresses nucleus-localized nCas9 (D10A) fused to PolI5M and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249067p-mCherryExpresses nucleus-localized mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249068pEvolvR-mCherry-enCas9(D10A)-PolI5MΔExpresses nucleus-localized enCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249069pEvolvR-nCas9(D10A)-PolI5MΔExpresses nucleus-localized nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249070pEvolvR-mCherry-SuperFi-nCas9(D10A)-PolI5MΔExpresses nucleus-localized SuperFi-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249071pEvolvR-mCherry-Sniper-nCas9(D10A)-PolI5MΔExpresses nucleus-localized Sniper-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249072pEvolvR-mCherry-LZ3-nCas9(D10A)-PolI5MΔExpresses nucleus-localized LZ3-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249073pEvolvR-mCherry-HSC1.2-nCas9(D10A)-PolI5MΔExpresses nucleus-localized HSC1.2-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249074pEvolvR-mCherry-SpRY-nCas9(D10A)-PolI5MΔExpresses nucleus-localized SpRY-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249075pEvolvR-mCherry-HF1-nCas9(D10A)-PolI5MΔExpresses nucleus-localized HF1-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249076pEvolvR-mCherry-Hypa-nCas9(D10A)-PolI5MΔExpresses nucleus-localized Hypa-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249077pEvolvR-mCherry-eNme2.C.NR-nCas9(D16A)-PolI5MΔExpresses nucleus-localized eNme2.C.NR-nCas9 (D16A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249078pEvolvR-mCherry-Slug-nCas9(D10A)-PolI5MΔExpresses nucleus-localized Slug-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.
249079pEvolvR-mCherry-NNG-Slug-nCas9(D10A)-PolI5MΔExpresses nucleus-localized NNG-Slug-nCas9 (D10A) fused to PolI5MΔ and mCherry using a CMV promoter. Includes a GFP cassette flanked by BsmbI cut sites to knock in and coexpress user-defined gRNAs.

Antibodies from Article