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Advancing knock-in approaches for robust genome editing in zebrafish

Rodriguez-Parks A, Beezley EG, Manna S, Silaban I, Almutawa SI, Cao S, Ahmed H, Guyer M, Baker S, Richards MP, Kang J
Biol Open 2026; bio.062472. doi: 10.1242/bio.062472 (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
200513pMC-ME_EGFP-pAMini circle middle entry plasmid for GFP-pA with flanking BsaI cut sites
200514pMC-ME_2a-EGFP-pA ORF-1 with ATGmini circle middle entry plasmid for 2a-eGFP-pA ORF1 (with ATG) with flanking BsaI cut sites
200515pMC-ME_2a-EGFP-pA ORF-2 with ATGmini circle middle entry plasmid for 2a-GFP-pA ORF 2 (with ATG) with flanking BsaI cut sites
200516pMC-ME_2a-EGFP-pA ORF-3 with ATGmini circle middle entry plasmid for 2a-GFP-pA-ORF 3 (with ATG) with flanking BsaI cut sites
200517pMC-ME_2a-EGFP-pA ORF-1 without ATGmini circle middle entry plasmid for 2a-eGFP-pA ORF1 (without ATG) with flanking BsaI cut sites
200518pMC-ME_2a-EGFP-pA ORF-2 without ATGmini circle middle entry plasmid for 2a-eGFP-pA ORF2 (without ATG) with flanking BsaI cut sites
200519pMC-ME_2a-EGFP-pA ORF-3 without ATGmini circle middle entry plasmid for 2a-eGFP-pA ORF3 (without ATG) with flanking BsaI cut sites
200520pMC-ME_memb-EGFP-pAMini circle middle entry plasmid for membrane GFP- pA with flanking BsaI cut sites
200521pMC-ME_mCherry-pAMini circle middle entry plasmid for mCherry- pA with flanking BsaI cut sites
200522pMC-ME_2a-mCherry-pA ORF-1 with ATGmini circle middle entry plasmid for 2a-mCherry-pA ORF 1 (with ATG) with flanking BsaI cut sites
200523pMC-ME_2a-mCherry-pA ORF-2 with ATGmini circle middle entry plasmid for 2a-mCherry-pA ORF 2 (with ATG) with flanking BsaI cut sites
200524pMC-ME_2a-mCherry-pA ORF-3 with ATGmini circle middle entry plasmid for 2a-mCherry-pA ORF 3 (with ATG) with flanking BsaI cut sites
200525pMC-ME_2a-mCherry-pA ORF-1 without ATGmini circle middle entry plasmid for 2a-mCherry-pA ORF 1 (without ATG) with flanking BsaI cut sites
200526pMC-ME_2a-mCherry-pA ORF-2 without ATGmini circle middle entry plasmid for 2a-mCherry-pA ORF 2 (without ATG) with flanking BsaI cut sites
200527pMC-ME_2a-mCherry-pA ORF-3 without ATGmini circle middle entry plasmid for 2a-mCherry-pA ORF 3 (without ATG) with flanking BsaI cut sites
200528pMC-ME_memb-mCherry-pAmini circle middle entry plasmid for membrane mCherry-pA with flanking BsaI cut sites
200529pMC-ME T2A-mCherry-P2A-NTRv2-pA ORF-1mini circle middle entry plasmid for T2A- mCherry- P2A- ntr version 2- pA (ORF 1) with flanking BsaI cut sites
200530pMC-ME T2A-mCherry-P2A-NTRv2-pA ORF-2mini circle middle entry plasmid for T2A- mCherry- P2A- ntr version 2- pA (ORF 2) with flanking BsaI cut sites
200531pMC-ME_T2A-mCherry-P2A-NTRv2-pA ORF-3mini circle middle entry plasmid for T2A- mCherry- P2A- ntr version 2- pA (ORF 3) with flanking BsaI cut sites
200532pMC-ME mCherry-P2A-NTRv2-pAmini circle middle entry plasmid for mCherry- P2A- ntr version 2- pA with flanking BsaI cut sites
200533pMC-ME_BFP-pAmini circle middle entry plasmid for BFP-pA with flanking BsaI cut sites
200534pMC-ME_2a-BFP-pA ORF-1 without ATGmini circle middle entry plasmid for 2a-BFP-pA ORF 1 (without ATG) with flanking BsaI cut sites
200535pMC-ME_2a-BFP-pA ORF-2 without ATGmini circle middle entry plasmid for 2a-BFP-pA ORF 2 (without ATG) with flanking BsaI cut sites
200536pMC-ME_2a-BFP-pA ORF-3 without ATGmini circle middle entry plasmid for 2a-BFP-pA ORF 3 (without ATG) with flanking BsaI cut sites
200537pMC-ME_memb-BFP-pAmini circle middle entry plasmid for membrane BFP-pA with flanking BsaI cut sites
200538pMC-ME_BFP-P2A-NTRv2-pAmini circle middle entry plasmid for BFP-P2A-NTR version 2-pA with flanking BsaI cut sites
200539pMC-ME_T2A-BFP-P2A-NTRv2-pA ORF-1mini circle middle entry plasmid for T2A- BFP- P2A- ntr version 2- pA (ORF 1) with flanking BsaI cut sites
200540pMC-ME_T2A-BFP-P2A-NTRv2-pA ORF-2mini circle middle entry plasmid for T2A- BFP- P2A- ntr version 2- pA (ORF 2) with flanking BsaI cut sites
200541pMC-ME_T2A-BFP-P2A-NTRv2-pA ORF-3mini circle middle entry plasmid for T2A- BFP- P2A- ntr version 2- pA (ORF 3) with flanking BsaI cut sites
200542pMC-ME_CreER-pAmini circle middle entry plasmid for CreER-pA with flanking BsaI cut sites
200543pMC-ME_CreER-pA-FRT-acry:venus-pA-FRTmini circle middle entry plasmid for CreER-pA-FRT-acry:venus-pA-FRT with flanking BsaI cut sites
200544pMC-ME_CreER-pA-FRT-acry:mCherry-pA-FRTmini circle middle entry plasmid for CreER-pA-FRT-acry: mCherry -pA-FRT with flanking BsaI cut sites
200545pMC-ME_P2A-CreER-pA ORF1 FRT-acry:mCherry-pA-FRTmini circle middle entry plasmid for P2A-CreER-pA (ORF 1) FRT-acry:mCherry-pA-FRT with flanking BsaI cut sites
200546pMC-ME_P2A-CreER-pA ORF2 FRT-acry:mCherry-pA-FRTmini circle middle entry plasmid for P2A-CreER-pA (ORF 2) FRT-acry:mCherry-pA-FRT with flanking BsaI cut sites
200547pMC-ME_P2A-CreER-pA ORF3 FRT-acry:mCherry-pA-FRTmini circle middle entry plasmid for P2A-CreER-pA (ORF 3) FRT-acry:mCherry-pA-FRT with flanking BsaI cut sites
200550pMC-Dest-BsaIMini circle destination vector with BsaI sites for Golden Gate Assembly
241150pMC-Dest-uni-gRNA5-BsaIDestination vector for mini-Golden subcloning platform; contains universal sgRNA at 5' end
241151EGFP-pAMiddle entry vector for mini-Golden subcloning platform; eGFP
241152memb-EGFP-pAMiddle entry vector for mini-Golden subcloning platform; eGFP
241153P2A-EGFP-pA-ORF-1 without ATGMiddle entry vector for mini-Golden subcloning platform; eGFP
241154P2A-EGFP-pA-ORF-2 without ATGMiddle entry vector for mini-Golden subcloning platform; eGFP
241155P2A-EGFP-pA-ORF-3 without ATGMiddle entry vector for mini-Golden subcloning platform; eGFP
241156mCherry-pAMiddle entry vector for mini-Golden subcloning platform; mCherry
241157memb-mCherry-pAMiddle entry vector for mini-Golden subcloning platform; mCherry
241158P2A-mCherry-pA-ORF-1 without ATGMiddle entry vector for mini-Golden subcloning platform; mCherry
241159P2A-mCherry-pA-ORF-2 without ATGMiddle entry vector for mini-Golden subcloning platform; mCherry
241160P2A-mCherry-pA-ORF-3 without ATGMiddle entry vector for mini-Golden subcloning platform; mCherry
241161memb-BFP-pAMiddle entry vector for mini-Golden subcloning platform; BFP
241162pMC-ME_2a-BFP-ORF-1 without ATGMiddle entry vector for mini-Golden subcloning platform; BFP
241163P2A-BFP-pA-ORF-2 without ATGMiddle entry vector for mini-Golden subcloning platform; BFP
241164P2A-BFP-pA-ORF-3 without ATGMiddle entry vector for mini-Golden subcloning platform; BFP
241165CreER-pA_FRT-acrymCherryMiddle entry vector for mini-Golden subcloning platform; creER; acry-mCherry
241166CreER-pA_FRT-acry-venusMiddle entry vector for mini-Golden subcloning platform; creER; acry-venus
241167P2A-CreER-pA-ORF1-FRT-acry mCherryMiddle entry vector for mini-Golden subcloning platform; creER
241168P2A-CreER-pA-ORF2-FRT-acry mCherryMiddle entry vector for mini-Golden subcloning platform; creER
241169P2A-CreER-pA-ORF3-FRT-acry mCherryMiddle entry vector for mini-Golden subcloning platform; creER
241170MCS_BsmBIMiddle entry vector for mini-Golden subcloning platform; MCS; BsmBI
241171pMid-TCGC-mNeongreenMiddle entry vector for mini-Golden subcloning platform; mNeongreen
241172linker-P2A-lifeact-mNeongreen-ORF-1Middle entry vector for mini-Golden subcloning platform; mNeongreen
241173linker-P2A-lifeact-mNeongreen-ORF-2Middle entry vector for mini-Golden subcloning platform; mNeongreen
241174linker-P2A-lifeact-mNeongreen-ORF-3Middle entry vector for mini-Golden subcloning platform; mNeongreen
241175memb-mNeongreenMiddle entry vector for mini-Golden subcloning platform; mNeongreen
241176linker-P2A-mNeongreen-ORF1Middle entry vector for mini-Golden subcloning platform; mNeongreen
241177linker-P2A-mNeongreen-ORF2Middle entry vector for mini-Golden subcloning platform; mNeongreen
241178linker-P2A-mNeongreen-ORF3Middle entry vector for mini-Golden subcloning platform; mNeongreen
241179FRT-acrymCherry-FRTMiddle entry vector for mini-Golden subcloning platform; acry-mCherry
241180linker-T2A-NTRv2-linker-P2A-mNeonGreen ORF1Middle entry vector for mini-Golden subcloning platform; NTRv2; mNeonGreen
241181linker-T2A-NTRv2-linker-P2A-mNeonGreen ORF2Middle entry vector for mini-Golden subcloning platform; NTRv2; mNeonGreen
241182linker-T2A-NTRv2-linker-P2A-mNeonGreen ORF3Middle entry vector for mini-Golden subcloning platform; NTRv2; mNeonGreen
241183linker-P2A-mem-mScarlet-ORF-1Middle entry vector for mini-Golden subcloning platform; mScarlet
241184linker-P2A-mem-mScarlet-ORF-2Middle entry vector for mini-Golden subcloning platform; mScarlet
241185linker-P2A-mem-mScarlet-ORF-3Middle entry vector for mini-Golden subcloning platform; mScarlet
241186SA-linker-P2A-mem-mScarlet-ORF-1Middle entry vector for mini-Golden subcloning platform; mScarlet
241187SA-linker-P2A-mem-mScarlet-ORF-2Middle entry vector for mini-Golden subcloning platform; mScarlet
241188SA-linker-P2A-mem-mScarlet-ORF-3Middle entry vector for mini-Golden subcloning platform; mScarlet
241189nzCas9n-GSG-linker-P2A-mNeongreenMiddle entry vector for mini-Golden subcloning platform; mNeongreen
2411903x-FLAG-nzdCas9-V5-APEX2n-GSG-linker-P2A-mNeongreenMiddle entry vector for mini-Golden subcloning platform; V5-APEX2n; mNeongreen
241191nAPEX2-V5-zdCas9n-3x-FLAG-GSG-linker-P2A-mNeongreenMiddle entry vector for mini-Golden subcloning platform; 3x-FLAG; mNeongreen
241192linker-T2A-NTRv2-linker-P2A-mNeonGreen-CAAX ORF1Middle entry vector for mini-Golden subcloning platform; NTRv2; mNeonGreen
241193linker-T2A-NTRv2-linker-P2A-mNeonGreen-CAAX ORF2Middle entry vector for mini-Golden subcloning platform; NTRv2; mNeonGreen
241194linker-T2A-NTRv2-linker-P2A-mNeonGreen-CAAX ORF3Middle entry vector for mini-Golden subcloning platform; NTRv2; mNeonGreen
241195mCherry-P2A-NTRv2-pAMiddle entry vector for mini-Golden subcloning platform; mCherry; NTR-v2
241196T2A-mCherry-P2A-NTRv2-pA-ORF-1Middle entry vector for mini-Golden subcloning platform; mCherry; NTR-v2
241197T2A-mCherry-P2A-NTRv2-pA-ORF-2Middle entry vector for mini-Golden subcloning platform; mCherry; NTR-v2
241198T2A-mCherry-P2A-NTRv2-pA-ORF-3Middle entry vector for mini-Golden subcloning platform; mCherry; NTR-v2
241199T2A-BFP-P2A-NTRv2-pA-ORF-1Middle entry vector for mini-Golden subcloning platform; BFP; NTR-v2
241200T2A-BFP-P2A-NTRv2-pA-ORF-2Middle entry vector for mini-Golden subcloning platform; BFP; NTR-v2
241201T2A-BFP-P2A-NTRv2-pA-ORF-3Middle entry vector for mini-Golden subcloning platform; BFP; NTR-v2
241202lifeact-mNeongreen-pAMiddle entry vector for mini-Golden subcloning platform; lifeact-mNeongreen
241203SA-TCGC-mNeongreenMiddle entry vector for mini-Golden subcloning platform; mNeongreen
241204SA-linker-P2A-lifeact-mNeongreen-ORF-1Middle entry vector for mini-Golden subcloning platform; lifeact-mNeongreen
241205SA-linker-P2A-lifeact-mNeongreen-ORF-2Middle entry vector for mini-Golden subcloning platform; lifeact-mNeongreen
241206SA-linker-P2A-lifeact-mNeongreen-ORF-3Middle entry vector for mini-Golden subcloning platform; lifeact-mNeongreen
241207SA-memb-mNeongreenMiddle entry vector for mini-Golden subcloning platform; mNeongreen
241208SA-linker-P2A-mNeongreen-ORF1Middle entry vector for mini-Golden subcloning platform; mNeongreen
241209SA-linker-P2A-mNeongreen-ORF2Middle entry vector for mini-Golden subcloning platform; mNeongreen
241210SA-linker-P2A-mNeongreen-ORF3Middle entry vector for mini-Golden subcloning platform; mNeongreen
241211SA-EGFP-pAMiddle entry vector for mini-Golden subcloning platform; eGFP
241212SA-mCherry-pAMiddle entry vector for mini-Golden subcloning platform; mCherry
241213pMC-dest-BsaI-cmlc2:mCherry
241214BsaI-MCS-cloning vector for golden gate

Antibodies from Article