A custom knock-in kit designed just for you
Are you performing an HDR knock-in experiment? Do you want to insert a custom sequence into your gene of interest? If your insert sequence is longer than 150 nucleotides*, we have developed a universal Edit-R HDR Plasmid Donor Kit that allows you to insert a custom sequence nearly anywhere in the genomes of over 30 different species. Use our HDR Donor Designer to design your own custom HDR kit that includes a plasmid backbone and primers specific to the genomic location and custom insert sequence to allow for a Gibson assembly of the HDR donor plasmid. One important part of the HDR plasmid is not included though, the custom insert. You must provide the DNA source for the custom insert sequence either as synthesized DNA, plasmid DNA, cDNA, PCR product, restriction digest fragment or genomic DNA. Do we have recommended sources for particular applications? So glad you asked!
Are you trying to affix a purification, epitope or reporter tag to your protein of interest?
A plasmid containing that particular DNA sequence can be an ideal source for amplification of that sequence. Please be aware that it is your responsibility to make sure that the terms and conditions of the source plasmid allows for its use in your knock-in experiment.
Do you want to insert a whole gene sequence into your cells?
The Mammalian Gene collection (MGC) contains over 100,000 human, mouse, rat and bovine genes. Isolated plasmid from MGC clones would allow the insertion of specific biologically derived gene content into a cell.
Would you like to insert a specific gene variant to recapitulate a rare, biologically-relevant genotype?
You can obtain this sequence by generating cDNA from a tissue or cell type. This would be a great way to amplify a sequence associated with a disease state to recapitulate the phenotype in a cell culture model. It has the advantage over an ORF or cDNA of replacing the endogenous gene, not expressing a foreign construct and endogenous gene at the same time.
Want to alter your gene (by add a binding or nuclease domain) to see how biological function is affected?
A domain sequence could be amplified from the genomic DNA of a tissue or cell.
Need to attach a red or green fluorescent reporter to your protein of interest?
No need to find a custom sequence. We offer HDR Donor kits for mKate2 and EGFP for this very purpose!
Whatever the source of the custom insert, our HDR Donor Designer helps you easily create and order primers to amplify the homology arms required for HDR. These primers are designed with linkers to perform a Gibson assembly of the entire HDR donor plasmid including the plasmid backbone, homology arms, and custom insert.
Have questions? Want to discuss experimental design and considerations? Please contact our Scientific support team!
* If the total insert length including homology arms is <150 nucleotides, then you’ll want to use a ssDNA donor in your experiments (learn more here).
Additional Resources
Edit-R Plasmid Donor Kits
- Kit for the assembly of a plasmid DNA EGFP, mKate2, or custom sequence HDR knock-in donor
HDR Donor Designer Workflow
- Quickly and easily design and order a donor template for precise CRISPR-Cas9 gene editing!