CRISPaint Gene Tagging Kit
(Kit #
1000000086
)
Depositing Lab: Veit Hornung
The CRISPaint (CRISPR-assisted insertion tagging) Kit, developed by the Hornung Lab, is a modular system for integrating large DNA fragments into user-defined genomic locations.
The CRISPaint system relies on a ligase-4 dependent mechanism (canonical NHEJ), instead of HDR, so there is no need for a site-specific donor template with homology arms. CRISPaint allows for tagging of endogenous proteins of interest. This kit includes several donor plasmids for tagging, including luciferase, fluorescent proteins, and small epitope tags. CRISPaint allows users to probe for endogenous protein expression, localization, and protein-protein interactions without the need for protein-specific antibodies.
This kit will be sent as bacterial glycerol stocks in 96-well plate format. It is recommended to grow the strains harboring the pCAS9 plasmids at 30°C, to limit potential plasmid recombination.
Original Publication
CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Schmid-Burgk JL, Höning K, Ebert TS, Hornung V. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. PubMed PMID: 27465542.
Description
CRISPaint is a three component system for tagging your gene of interest. To use this system, you need:
- to construct a gRNA plasmid targeting your gene of interest
- a plasmid to specify the insertion reading frame ( in this kit )
- a donor plasmid containing the tag itself ( in this kit )
DNA cassettes provided with this kit include luciferase, various fluorescent proteins, and epitope tags.
Immunofluorescence of cells with tagged gene expression using CRISPaint. Clockwise image description:
- Top-left: Nucleoli
- Top-right: Microtubuli
- Bottom-right: Chromatin
- Bottom-left: Mitochondria
Reference Figure 4 in Schmid-Burgk et al, Nature Communications doi:10.1038/ncomms12338
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The CRISPaint Kit was a gift from Veit Hornung (Addgene kit # 1000000086).”
For your Reference section:
CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Schmid-Burgk JL, Höning K, Ebert TS, Hornung V. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. PubMed PMID: 27465542.
CRISPaint Gene Tagging Kit - #1000000086
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Ampicillin |