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Marker-free co-selection for successive rounds of prime editing in human cells.

Levesque S, Mayorga D, Fiset JP, Goupil C, Duringer A, Loiselle A, Bouchard E, Agudelo D, Doyon Y
Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
173199ATP1A1_G4_Q118R_Dual_pegRNACoselection for prime editing in human cells. Vector for tandem expression of ATP1A1 Q118R-G4 pegRNA with a user-specified pegRNA from two independent U6 promoters. pU6-pegRNA-GG-acceptor-like plasmid
173200ATP1A1_G6_T804N_Dual_pegRNACoselection for prime editing in human cells. Vector for tandem expression of ATP1A1 T804N-G6 pegRNA with a user-specified pegRNA from two independent U6 promoters. pU6-pegRNA-GG-acceptor-like plasmid
173201ATP1A1_G2_Dual_sgRNACoselection for ABE or NHEJ in human cells. Vector for tandem expression of ATP1A1 G2 sgRNA with a user-specified sgRNA from two independent U6 promoters. Cloning of oligos using BbsI sites.
173202ATP1A1_G3_Dual_sgRNACoselection for PE3 or HDR in human cells. Vector for tandem expression of ATP1A1 G3 sgRNA with a user-specified PE3 nick sgRNA from two independent U6 promoters. Cloning of oligos using BbsI sites.
173203eSpCas9(1.1)_No_FLAG_ATP1A1-G7Expresses the ATP1A1 G7 sgRNA in combination with FLAGless eSpCas9(1.1) to target ATP1A1 intron 17. pX330-like plasmid.
173204pX330_ATP1A1_G7Expresses the ATP1A1 G7 sgRNA in combination with SpCas9 to target ATP1A1 intron 17.
173205pX330_ATP1A1_G3_Dual_sgRNACoselection for HDR in human cells. Vector for dual expression of ATP1A1 G3 sgRNA in combination with a user-specified sgRNA from two independent U6 promoters. Cloning of oligos using BbsI sites.
173206pX330_ATP1A1_G7_Dual_sgRNACoselection for HDR in human cells. Vector for dual expression of ATP1A1 G7 sgRNA in combination with a user-specified sgRNA from two independent U6 promoters. Cloning of oligos using BbsI sites.
173207ATP1A1_T804N_hPGK1_mScarlet-I_DonorHDR donor plasmid to introduce the T804N mutation conferring cellular resistance to ouabain and the mScarlet-I cassette to ATP1A1 intron 17.
173208ATP1A1_T804N_hPGK1_mScarlet-I-TOSI_DonorHDR donor plasmid to introduce the T804N mutation conferring cellular resistance to ouabain and the mScarlet-I mTOR signaling indicator (TOSI) cassette to ATP1A1 intron 17.
173209ATP1A1_G4_Q118R_MTOR_E2419K_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and MTOR-E2419K pegRNAs from two independent U6 promoters.
173211ATP1A1_G3_MTOR_Nick_Exon-53_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G3 and MTOR Nick exon-53 sgRNAs from two independent U6 promoters.
173212ATP1A1_G4_Q118R_RUNX1_+4_T_to_G_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and RUNX1 +4 T to G pegRNAs from two independent U6 promoters.
173213ATP1A1_G4_Q118R_RUNX1_+1_ATGins_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and RUNX1 +1 ATG insertion pegRNAs from two independent U6 promoters.
173214ATP1A1_G3_RUNX1_Nick-38_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G3 and RUNX1 nick-38 sgRNAs from two independent U6 promoters.
178087AAVS1_Puro_hPGK1_mScarlet-I-TOSI_DonorHDR donor plasmid to introduce the mScarlet-I mTOR signaling indicator (TOSI) cassette to AAVS1 using AAVS1 T2 sgRNA.
178088AAVS1_Puro_hPGK1_EGFP_DonorHDR donor plasmid to introduce the EGFP cassette to AAVS1 using AAVS1 T2 sgRNA.
178089AAVS1_Puro_hPGK1_EBFP_DonorHDR donor plasmid to introduce the EBFP (T65S/F145Y) cassette to AAVS1 using AAVS1 T2 sgRNA.
178090eSpCas9(1.1)_No_FLAG_LMNA_G2Expresses the LMNA G2 sgRNA in combination with FLAGless eSpCas9(1.1). This vector can be used in combination with LMNA_mScarlet-I_Donor to tag LMNA with mScarlet-I. pX330-like plasmid.
178091eSpCas9(1.1)_No_FLAG_NPM1_G6Expresses the NPM1 G6 sgRNA in combination with FLAGless eSpCas9(1.1). This vector can be used in combination with NPM1_mNeonGreen_Donor to tag NPM1 with mNeonGreen. pX330-like plasmid.
178092LMNA_mScarlet-I_DonorHDR donor plasmid to tag LMNA with mScarlet-I using eSpCas9(1.1)_No_FLAG_LMNA_G2.
178093NPM1_mNeonGreen_DonorHDR donor plasmid to tag NPM1 with mNeonGreen using eSpCas9(1.1)_No_FLAG_NPM1_G6.
178094ATP1A1_G3_EBFP_Nick_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G3 and EBFP Nick sgRNAs from two independent U6 promoters.
178095ATP1A1_G3_EGFP_Nick_Dual_sgRNAControl vector for coselection for PE3b in human cells. Tandem expression of ATP1A1 G3 and EGFP Nick sgRNAs from two independent U6 promoters.
178096ATP1A1_G3_MTOR_Nick_Exon-44_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G3 and MTOR Nick exon-44 sgRNAs from two independent U6 promoters.
178099ATP1A1_G3_MTOR_Nick_Intron-45_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G3 and MTOR Nick intron-45 sgRNAs from two independent U6 promoters.
178100ATP1A1_G4_Q118R_EBFP_to_EGFP_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and EBFP_To_EGFP pegRNAs from two independent U6 promoters.
178102ATP1A1_G4_Q118R_MTOR_F2108L_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and MTOR-F2108L pegRNAs from two independent U6 promoters.
178103ATP1A1_G4_Q118R_MTOR_I2017T_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 and MTOR-I2017T pegRNAs from two independent U6 promoters.
178104ATP1A1_G8_Dual_sgRNACoselection for PE3 in human cells. Vector for tandem expression of ATP1A1 G8 sgRNA with a user-specified PE3 nick sgRNA from two independent U6 promoters. Cloning of oligos using BbsI sites.
178105ATP1A1_G8_MTOR_Nick_Exon-44_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G8 and MTOR Nick exon-44 sgRNAs from two independent U6 promoters.
178106ATP1A1_G8_MTOR_Nick_Intron-45_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G8 and MTOR Nick intron-45 sgRNAs from two independent U6 promoters.
178107ATP1A1_G8_MTOR_Nick_Exon-53_Dual_sgRNAControl vector for coselection for PE3 in human cells. Tandem expression of ATP1A1 G8 and MTOR Nick exon-53 sgRNAs from two independent U6 promoters.
178108ATP1A1_G6_T804N_MTOR_I2017T_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 T804N-G6 and MTOR-I2017T pegRNAs from two independent U6 promoters.
178109ATP1A1_G6_T804N_MTOR_F2108L_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 T804N-G6 and MTOR-F2108L pegRNAs from two independent U6 promoters.
178110ATP1A1_G6_T804N_MTOR_E2419K_Dual_pegRNAControl vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 T804N-G6 and MTOR-E2419K pegRNAs from two independent U6 promoters.
187454ATP1A1_G4_Q118R_Std_Dual_epegRNA_tevopreQ1Coselection for prime editing in human cells. Vector for tandem expression of ATP1A1 Q118R-G4 pegRNA with a user-specified epegRNA. pU6-tevopreq1-GG-acceptor-like plasmid
187455ATP1A1_T804N_hPGK1_EBFP_DonorHDR donor plasmid to introduce the T804N mutation conferring cellular resistance to ouabain and the EBFP cassette to ATP1A1 intron 17.
187456ATP1A1_G4_Q118R_v2_Dual_epegRNA_tevopreQ1Coselection for prime editing in human cells. Vector for tandem expression of ATP1A1 Q118R-G4_v2 optimized epegRNA with a user-specified epegRNA. pU6-tevopreq1-GG-acceptor-like plasmid
187457ATP1A1_G6_T804N_Std_Dual_epegRNA_tevopreQ1Coselection for prime editing in human cells. Vector for tandem expression of ATP1A1 T804N-G6 pegRNA with a user-specified epegRNA. pU6-tevopreq1-GG-acceptor-like plasmid
187458ATP1A1_G6_T804N_v1_Dual_epegRNA_tevopreQ1Coselection for prime editing in human cells. Vector for tandem expression of ATP1A1 T804N-G6 epegRNA (tevopreQ1) with a user-specified epegRNA. pU6-tevopreq1-GG-acceptor-like plasmid
187459ATP1A1_G4_Q118R_Std_BFP-to-EGFP_Dual_epegRNA_tevopreQ1Control vector for coselection for prime editing in human cells. Tandem expression of ATP1A1 Q118R-G4 pegRNA and EBFP_To_EGFP epegRNA (tevopreq1) from two independent U6 promoters.

Antibodies from Article