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Promoter strength and position govern promoter competition through transcript-dependent insulation.

Koska M, Nagano M, Swigut T, Boettiger AN, Hansen AS, Wysocka J
Nat Genet. 2026 Jul 20. doi: 10.1038/s41588-026-02691-y. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
253414pB_hU6_Cas9-t2a-PuropiggyBac integration and stable expression of sgRNA and SpCas9. Based on plasmid px459 (Addgene #62988)
253415pB_hU6_dCas9-t2a-PuropiggyBac integration and stable expression of guide and dCas9.
253416CARGO_Ef1a-6mer6-mer gRNA array targeting Sp dCas9/Cas9 to the Ef1a promoter
253417CARGO_mTagBFP2-6mer6-mer gRNA array targeting Sp dCas9/Cas9 to the mTagBFP2 transgene with intron
253418CARGO_Neutral-6mer6-mer gRNA array targeting Sp dCas9/Cas9 to the Neutral promoter (Ecoli uraA CDS)
253419pGEM-T-hU6-spSL-CARGOConstantEmpty backbone with CARGO constant region
253420pK335_CARGO-NoReporterBackbone for CARGO cloning, with U6 promoter, gRNA scaffold and Kanamycin resistence, without flurophore
253421Rad21-C-term-dTagPuroC-terminal tagging of mouse Rad21 with FKBP and Puromycin
253422SCR-CTCFmutatedintroduces CTCF mutations and a selection casette into the core SCR
253423pDonorRMCE-For-FRT-MCS-mTagBFP2wintron-SV40pA-FRT3Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with MCS for promoter integrations
253424pDonorRMCE-Rev-FRT3-MCS-mTagBFP2wintron-SV40pA-FRTDonor plasmid for FLP/FRT mediated recombination into landing pad in the reverse direction with MCS for promoter integrations
253425pDonorRMCE-For-FRT-MCS-mTagBFP2wintron-noSV40pA-FRT3Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with MCS for promoter integrations, without an SV40 polyA signal after reporter
253426pDonorRMCE-Rev-FRT3-MCS-mTagBFP2wintron-noSV40pA-FRTDonor plasmid for FLP/FRT mediated recombination into landing pad in the reverse direction with MCS for promoter integrations, without an SV40 polyA signal after reporter
253427pDonorRMCE-For-Rpl13a-CTCFDeleted-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with Rpl13a promoter (CTCF sites are deleted)
253428pDonorRMCE-For-Rpl13a-CTCFScrambled-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with Rpl13a promoter (CTCF sites are mutated)
253429pDonorRMCE-For-dBx-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dBx
253430pDonorRMCE-For-dPst-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dPst
253431pDonorRMCE-For-dSac-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dSac
253432pDonorRMCE-For-dP2-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dP2
253433pDonorRMCE-For-dP4-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dP4
253434pDonorRMCE-For-dl1-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dl1
253435pDonorRMCE-For-dl2-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dl2
253436pDonorRMCE-For-dl3-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dl3
253437pDonorRMCE-For-dl6-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dl6
253438pDonorRMCE-For-dl8-mTagBFP2Donor plasmid for FLP/FRT mediated recombination into landing pad in the forward direction with mutant Ef1a promoter - dl8
253439pDonorRMCE-For-Ef1a-mTagBFP2-luciferaseModified donor plasmid for FLP/FRT RMCE in forward direction with longer reporter (mTagBFP2+luciferase) and Ef1a promoter
253440pDonorRMCE-Rev-Ef1a-mTagBFP2-luciferaseModified donor plasmid for FLP/FRT RMCE in reverse direction with longer reporter (mTagBFP2+luciferase) and Ef1a promoter
253441pDonorRMCE-For-Neutral-mTagBFP2-luciferaseModified donor plasmid for FLP/FRT RMCE in forward direction with longer reporter (mTagBFP2+luciferase) and Neutral promoter
253442pDonorRMCE-Rev-Neutral-mTagBFP2-luciferaseModified donor plasmid for FLP/FRT RMCE in reverse direction with longer reporter (mTagBFP2+luciferase) and Neutral promoter

Antibodies from Article