PLL5.0-Anillin ShRNA-Cherry-AnillinA740D, E758K
PurposeLentiviral expression of shRNA targeting Anillin + reconstitution of Anillin A740D,E758K
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||187273||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 7593
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Gene/Insert nameAnillin ,hsAnillin, Entrez Gene ANLN (a.k.a. FSGS8, Scraps, scra)
SpeciesH. sapiens (human)
Insert Size (bp)3324
MutationAnillin A740D, E758K
Entrez GeneANLN (a.k.a. FSGS8, Scraps, scra)
- Promoter U6, CMV
/ Fusion Protein
- mCherry (N terminal on insert)
- Cloning method Gibson Cloning
- 5′ sequencing primer mU6 Forward Primer,mCherry-F
- 3′ sequencing primer WPRE-R (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Rescue with reconstituted -Anillin A740D,E758K.
5' cloning site: SacII (not destroyed), 3' cloning site: Sbf1 (destroyed).
With regard to all the Anillin sequences, I chose to number these residues as per the canonical sequence which is the full-length isoform. We have been using a smaller isoform, that lacks aa 508-544 as compared to the canonical isoform. The article that identified the Rho binding region on Anillin (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4449299/) followed the residue numbers of the canonical isoform to describe the mutations a740D, E758K (ref figure 2 in the article). Therefore, I chose to follow similar nomenclature when I generated the AHPH-DM for Rashmi's NCB paper. To harmonize the nomenclature across constructs, I continued to number the residues in all the GFP/Cherry Anillin constructs as per the canonical sequence.
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:PLL5.0-Anillin ShRNA-Cherry-AnillinA740D, E758K was a gift from Alpha Yap (Addgene plasmid # 187273 ; http://n2t.net/addgene:187273 ; RRID:Addgene_187273)
For your References section:Anillin Promotes Cell Contractility by Cyclic Resetting of RhoA Residence Kinetics. Budnar S, Husain KB, Gomez GA, Naghibosadat M, Varma A, Verma S, Hamilton NA, Morris RG, Yap AS. Dev Cell. 2019 Jun 17;49(6):894-906.e12. doi: 10.1016/j.devcel.2019.04.031. Epub 2019 May 16. 10.1016/j.devcel.2019.04.031 PubMed 31105010