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Genome and epigenome engineering CRISPR toolkit for in vivo modulation of cis-regulatory interactions and gene expression in the chicken embryo.
Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, Sauka-Spengler T
Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 10.1242/dev.160333.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
92357pCAG MCP-VP64CAG-driven ubiquitous expression of MCP-VP64. MCP is bacteriophage MS2 coat protein, VP64 is transcriptional activation domain. Part of SAM-mediated gene activation system in chicken embryos.
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92358pCAG Cas9-2A-CitrineCAG-driven ubiquitous expression of Cas9. Contains 2A-Citrine reporter. For CRISPR mediated gene knockouts in chicken embryos.
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92359pcU6_3 sgRNAChicken-specific U6 sgRNA expression mini-vector, harbouring chick U6_3 pol III promoter.
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92361pX330a dCas9-KRABModified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to KRAB repressor domain. For targeted enhancer inactivation in chicken embryos.
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92362pX330a dCas9-LSD1Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to LSD1. For targeted enhancer demethylation in chicken embryos.
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92363pX330a dCas9-VP64Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to VP64 transcriptional activation domain. For targeted gene activation in chicken embryos.
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92393pTK Cas9-2A-CitrineEnhancer/reporter plasmid for tissue-specific expression of Cas9 with 2A-Citrine reporter. Contains BsmBI-flanked LacZ cloning cassette for rapid GoldenGate-based cloning of specific enhancers.
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92394pcU6_3 MS2 sgRNAChick-specific U6 sgRNA expression mini-vector, harbouring chick U6_3 pol III promoter, with tracrRNA scaffold containing stem loops allowing binding of bacteriophage MS2 coat protein MCP
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92395pcU6_1 sgRNAChicken-specific U6 sgRNA expression mini-vector, harbouring chick U6_1 pol III promoter.
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92396pCAG dCas9-KRAB-2A-EGFPCAG-driven ubiquitous expression of catalytically inactive Cas9 fused to KRAB repressor domain. Contains 2A-EGFP reporter. For targeted enhancer inactivation in chicken embryos.
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92397pCI Cas9 H2B-RFPCAG-driven ubiquitous expression of Cas9 with Histone2B-RFP reporter controlled by IRES
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92398pCI H2B-RFPUbiquitous expression plasmid, contains CAG promoter (CMV immediate early enhancer and chicken beta actin promoter), MCS and IRES controlled Histone2B-mRFP1 reporter.
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92399pCI H2B-GFPUbiquitous expression plasmid, contains CAG promoter (CMV immediate early enhancer and chicken beta actin promoter), MCS and IRES controlled Histone2B-EGFP reporter.
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