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Site-specific DNA insertion into the human genome with engineered recombinases.

Fanton A, Bartie LJ, Martins JQ, Tran VQ, Goudy L, Kernick C, Durrant MG, Wei J, Armour-Garb Z, Pawluk A, Konermann S, Marson A, Gilbert LA, Roth TL, Hsu PD
Nat Biotechnol. 2025 Nov 6. doi: 10.1038/s41587-025-02895-3. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
247151pEf1a-Dn29-P2A-GFPMammalian expression of a human codon optimized wildtype Dn29 recombinase
247152pEf1a-superDn29-P2A-GFPMammalian expression of a human codon optimized engineered superDn29 recombinase (highest efficiency variant)
247153pEf1a-goldDn29-P2A-GFPMammalian expression of a human codon optimized engineered goldDn29 recombinase (efficient and specific variant)
247154pEf1a-hifiDn29-P2A-GFPMammalian expression of a human codon optimized engineered hifiDn29 recombinase (highest specificity variant)
247155pEf1a-Dn29-dCas9-P2A-GFPMammalian expression of a human codon optimized wildtype Dn29 recombinase fused to dCas9 and gRNA expression cassette for targeting attH1 with H1-g3
247156pEf1a-superDn29-dCas9-P2A-GFPMammalian expression of a human codon optimized engineered superDn29 recombinase fused to dCas9 and gRNA expression cassette for targeting attH1 with H1-g3
247157pEf1a-goldDn29-dCas9-P2A-GFPMammalian expression of a human codon optimized engineered goldDn29 recombinase fused to dCas9 and gRNA expression cassette for targeting attH1 with H1-g3
247158pEf1a-hifiDn29-dCas9-P2A-GFPMammalian expression of a human codon optimized engineered hifiDn29 recombinase fused to dCas9 and gRNA expression cassette for targeting attH1 with H1-g3
247159pDonor_attP-Ef1a-Puro-P2A-mCherryattP donor plasmid expressing puromycin resistance and mCherry
247160pDonor_e-attP-H1-g3-Ef1a-Puro-P2A-mCherryEngineered e-attP donor plasmid expressing puromycin resistance and mCherry with donor binding sgRNA H1-g3
247161pIVT_Dn29-P2A-GFPIn vitro transcription of human codon optimized wildtype Dn29-P2A-GFP
247162pIVT_superDn29-P2A-GFPIn vitro transcription of human codon optimized engineered superDn29-P2A-GFP
247163pIVT_goldDn29-P2A-GFPIn vitro transcription of human codon optimized engineered goldDn29-P2A-GFP
247164pIVT_hifiDn29-P2A-GFPIn vitro transcription of human codon optimized engineered hifiDn29-P2A-GFP
247165pIVT_Dn29-dCas9-P2A-GFPIn vitro transcription of human codon optimized fusion of Dn29-dCas9-P2A-GFP
247166pIVT_superDn29-dCas9-P2A-GFPIn vitro transcription of human codon optimized fusion of superDn29-dCas9-P2A-GFP
247167pIVT_goldDn29-dCas9-P2A-GFPIn vitro transcription of human codon optimized fusion of goldDn29-dCas9-P2A-GFP
247168pIVT_hifiDn29-dCas9-P2A-GFPIn vitro transcription of human codon optimized fusion of hifiDn29-dCas9-P2A-GFP
247169pU6_H1-g3U6 driven gRNA expression of H1-g3 for targeting Dn29 attH1

Antibodies from Article