PurposeModified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to KRAB repressor domain. For targeted enhancer inactivation in chicken embryos.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||92361||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4332
- Total vector size (bp) 8731
Vector typeMammalian Expression, CRISPR
Growth in Bacteria
Copy numberHigh Copy
Insert Size (bp)4399
- Promoter CAG
- Cloning method Gibson Cloning
- 5′ sequencing primer tctgactgaccgcgttactc
- 3′ sequencing primer aggaaaggacagtgggagtg (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byCas9m4-VP64 was amplified from (Addgene #47319) and cloned into pX330 (Addgene #42230). VP64 was then removed by EcoRI digest and KRAB was inserted by In-fusion of a synthetic gBlock containing the KRAB sequence into the EcoRI-linearised pX330 vector.
Terms and Licenses
CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Mali P, Aach J, Stranges PB, Esvelt KM, Moosburner M, Kosuri S, Yang L, Church GM. Nat Biotechnol. 2013 Aug 1. doi: 10.1038/nbt.2675. 10.1038/nbt.2675 PubMed 23907171
Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F. Science. 2013 Jan 3. 10.1126/science.1231143 PubMed 23287718
Highly specific epigenome editing by CRISPR-Cas9 repressors for silencing of distal regulatory elements. Thakore PI, D'Ippolito AM, Song L, Safi A, Shivakumar NK, Kabadi AM, Reddy TE, Crawford GE, Gersbach CA. Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 2015 Oct 26. 10.1038/nmeth.3630 PubMed 26501517
Please visit https://www.biorxiv.org/content/early/2017/05/08/135525 for bioRxiv preprint.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pX330a dCas9-KRAB was a gift from Tatjana Sauka-Spengler (Addgene plasmid # 92361 ; http://n2t.net/addgene:92361 ; RRID:Addgene_92361)
For your References section: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. 10.1242/dev.160333 PubMed 29386245