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Reduced Cas9 transgene silencing by incorporation of intron sequences.

Arana S, Du PP, Vaughan-Jackson A, Enright N, Spees K, Valbuena R, Garcia CA, Nguyen T, Venida A, Seczynska M, Bintu L, Lehner PJ, Prolo LM, Bassik MC
Nat Commun. 2025 Nov 27;16(1):10656. doi: 10.1038/s41467-025-65669-0. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
249132pSA071 Cas9-BFPOverexpression of SpCas9-BFP and blasticidin resistance gene.
249133pSA072 Cas9-HBB_IVS2-BFPOverexpression of SpCas9-BFP with HBB IVS2 intron after Cas9 coding sequence and blasticidin resistance gene. Contains Cp36 recombination site.
249134pSA073 HBB_IVS2-Cas9-BFPOverexpression of SpCas9-BFP with HBB IVS2 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249135pSA074 Cas9-BFP/HBB_IVS2/BFPOverexpression of SpCas9-BFP with HBB IVS2 intron within BFP coding sequence and blasticidin resistance gene. Contains Cp36 recombination site.
249136pSA075 Cas9-BFP-HBB_IVS2Overexpression of SpCas9-BFP with HBB IVS2 intron in 3' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249137pSA076 Cas9/HBB_IVS2/Cas9-BFPOverexpression of SpCas9-BFP with HBB IVS2 intron within Cas9 coding sequence and blasticidin resistance gene. Contains Cp36 recombination site.
249138pSA081 BFPOverexpression of BFP. Contains Cp36 recombination site.
249139pSA089 AARS1_IVS11-Cas9-BFPOverexpression of SpCas9-BFP with AARS IVS11 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249140pSA090 EIF4A3_IVS2-Cas9-BFPOverexpression of SpCas9-BFP with EIF4A3 IVS2 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249141pSA091 PSMD14_IVS2-Cas9-BFPOverexpression of SpCas9-BFP with PSMD14 IVS2 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249142pSA092 CCT3_IVS13-Cas9-BFPOverexpression of SpCas9-BFP with CCT3 IVS13 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249143pSA093 PFDN2_IVS3-Cas9-BFPOverexpression of SpCas9-BFP with PFDN2 IVS3 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249144pSA094 UBA1_IVS20-Cas9-BFPOverexpression of SpCas9-BFP with UBA1 IVS20 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249145pSA095 EEF2_IVS9-Cas9-BFPOverexpression of SpCas9-BFP with EEF2 IVS9 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249146pSA096 PSMD4_IVS6-Cas9-BFPOverexpression of SpCas9-BFP with PSMD4 IVS6 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249147pSA097 HNRNPK_IVS3-Cas9-BFPOverexpression of SpCas9-BFP with HNRNPK IVS3 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249148pSA098 RPL18A_IVS4-Cas9-BFPOverexpression of SpCas9-BFP with RPL18A IVS4 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249149pSA099 PGK1_IVS3-Cas9-BFPOverexpression of SpCas9-BFP with PGK1 IVS3 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249150pSA100 RPS12_IVS1-Cas9-BFPOverexpression of SpCas9-BFP with RPS12 IVS1 intron in 5' UTR and blasticidin resistance gene. Contains Cp36 recombination site.
249151pSA127 A2UCOE-Cas9-BFPOverexpression of SpCas9-BFP and blasticidin resistance gene. Contains minimal A2UCOE and Cp36 recombination site.
249152pSA128 A2UCOE-HBB_IVS2-Cas9-BFPOverexpression of SpCas9-BFP with HBB IVS2 intron in 5' UTR and blasticidin resistance gene. Contains minimal A2UCOE and Cp36 recombination site.
249153pSA143 Cas9-BFP with core cHS4 insulatorsOverexpression of SpCas9-BFP and blasticidin resistance gene. Flanked by cHS4 core insulators. Contains Cp36 recombination site.
249154pSA144 HBB_IVS2-Cas9-BFP with core cHS4 insulatorsOverexpression of SpCas9-BFP with HBB IVS2 intron in 5' UTR and blasticidin resistance gene. Flanked by cHS4 core insulators. Contains Cp36 recombination site.
249155pPD231 PPH-T2A-dCas9-NFZ-P2A-BFPOverexpression of PPH-T2A-dCas9-NFZ-P2A-BFP and blasticidin resistance gene. Contains Cp36 recombination site.
249156pPD292 PPH/HBB_IVS2/-T2A-dCas9-NFZ-P2A-BFPOverexpression of PPH-T2A-dCas9-NFZ-P2A-BFP with HBB IVS2 intron in PPH coding sequence and blasticidin resistance gene. Contains Cp36 recombination site.
249157pPD684 EnAsCas12a-BFPOverexpression of EnAsCas12a-BFP with blasticidin resistance gene. Contains Cp36 recombination site.
249158pPD685 HBB_IVS2-EnAsCas12a-BFPOverexpression of EnAsCas12a-BFP with HBB IVS2 intron in 5' UTR with blasticidin resistance gene. Contains Cp36 recombination site.
249159pPD686 A2UCOE-EnAsCas12a-BFPOverexpression of EnAsCas12a-BFP with blasticidin resistance gene. Contains minimal A2UCOE and Cp36 recombination site.
249160pPD687 A2UCOE-HBB_IVS2-EnAsCas12a-BFPOverexpression of EnAsCas12a-BFP with HBB IVS2 intron in 5' UTR with blasticidin resistance gene. Contains minimal A2UCOE and Cp36 recombination site.

Antibodies from Article