Custom guide cloning
Design your own guides with our tutorial and we'll clone them into lentivectors for you.
gRNA cloning service

We now offer a CRISPR/Cas9 cloning service that includes the annealing and cloning of your custom oligos into the single guide RNA scaffold of lentiviral vectors. You can choose between two different lentivectors: lentiCRISPRv2, which system allows the delivery of the desired single guide RNA (sgRNA) and Cas9 nuclease in mammalian cells, and lentiGuide-Puro, which allows the expression of the chimeric guide RNA only. It does not contain Cas9.

Controls: Non-targeting control constructs are available with same gRNA sequences as CRISPR19 and CRISPR20 controls in the Sanger library.

We can also obtain lentiCRISPRv2GFP upon request.

 

Guide design

Please refer to our step-by-step guide (pdf) on how to design gRNA sequences using the Benchling online program as a tool. It is also possible to use a different tool to design the guides, but keep in mind that there are specific sequences to incorporate in the oligo for the cloning to work.

Oligonucleotide ordering tips: Standard de-salted oligos (usually the most inexpensive synthesis) are sufficient for cloning. If not already resuspended, dilute each oligo to 100 μM in DNAse-free water.

Prices
  1st item in batch Rest
University of Helsinki 120 € 60 €
Biocenter Finland 140 € 70 €
External 435 € 110 €

A batch is defined as samples of one customer processed at the same time. Control constructs cost 20 € /miniprep. 

Citation

If you use a clone from the GBU in your publication, please acknowledge us in the following way:

“gRNA constructs were generated by the Genome Biology Unit supported by HiLIFE and the Faculty of Medicine, University of Helsinki, and Biocenter Finland"

Please also cite creators of the vector backbone of your construct. Details can be found on Addgene site for each plasmid:
lentiCRISPR v2
lentiGuide-Puro