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
92357	pCAG MCP-VP64	CAG-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.
92358	pCAG Cas9-2A-Citrine	CAG-driven ubiquitous expression of Cas9. Contains 2A-Citrine reporter. For CRISPR mediated gene knockouts in chicken embryos.
92359	pcU6_3 sgRNA	Chicken-specific U6 sgRNA expression mini-vector, harbouring chick U6_3 pol III promoter.
92361	pX330a dCas9-KRAB	Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to KRAB repressor domain. For targeted enhancer inactivation in chicken embryos.
92362	pX330a dCas9-LSD1	Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to LSD1. For targeted enhancer demethylation in chicken embryos.
92363	pX330a dCas9-VP64	Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to VP64 transcriptional activation domain. For targeted gene activation in chicken embryos.
92393	pTK Cas9-2A-Citrine	Enhancer/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.
92394	pcU6_3 MS2 sgRNA	Chick-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
92395	pcU6_1 sgRNA	Chicken-specific U6 sgRNA expression mini-vector, harbouring chick U6_1 pol III promoter.
92396	pCAG dCas9-KRAB-2A-EGFP	CAG-driven ubiquitous expression of catalytically inactive Cas9 fused to KRAB repressor domain. Contains 2A-EGFP reporter. For targeted enhancer inactivation in chicken embryos.
92397	pCI Cas9 H2B-RFP	CAG-driven ubiquitous expression of Cas9 with Histone2B-RFP reporter controlled by IRES
92398	pCI H2B-RFP	Ubiquitous expression plasmid, contains CAG promoter (CMV immediate early enhancer and chicken beta actin promoter), MCS and IRES controlled Histone2B-mRFP1 reporter.
92399	pCI H2B-GFP	Ubiquitous expression plasmid, contains CAG promoter (CMV immediate early enhancer and chicken beta actin promoter), MCS and IRES controlled Histone2B-EGFP reporter.