Custom Genome EngineeringOur partner SBI will design and clone gRNA as well as homology arms into any SBI gRNA and HR donor plasmid for your knock-out, knock-in, tagging, or single nucleotide modification genome engineering projects.
On top of providing customized gRNA and HR donor constructs SBI can engineer your genome edited cell line of choice.
The services can be ordered in a modular way as follows:
Custom gRNA Cas9 Design and Cloning
SBI will design and clone a gRNA against a target locus into any SBI SmartNuclease or SmartNickase vector (or customer provided gRNA cloning vector) for your desired knock-out, knock-in, tagging, or single nucleotide modification genome engineering project.
Turn-around time is about 10 days after customer approval of gRNA sequence design.
Wild-type or mutant, which Cas9 is right for you?
Wild-type PrecisionX Cas9 SmartNuclease generates double-strand breaks (DSBs), and can be used with an HR donor plasmid for efficient, targeted genome engineering. The mutant PrecisionX Cas9 SmartNickase (D10A mutation) creates nicks in genomic DNA instead of DSBs. Creating nicks favors the higher-fidelity homologous recombination process over non-homologous end joining (NHEJ), with paired nicking shown to reduce offtarget activity by 50- to 1,500 fold in cell lines, and to facilitate gene knockout in mice without losing on-target cleavage efficiency. SBI also offers a null PrecisionX Cas9 SmartNuclease (D10A and H840A mutations) for experimental controls. This mutant has both nuclease and nickase activities completely inactivated.
See link below for further information about SBI´s gRNA vector selection.
Custom HR Donor Design and Cloning
SBI will design and clone homology arms into any SBI HR donor plasmid for knock-out, knock-in, tagging, or single nucleotide modification genome engineering projects. When a custom HR donor plasmid design is ordered together with custom gRNA design and cloning, the donor vector will not contain full gRNA sequences in the homology arms to ensure full compatibility with gRNA. Turn-around time is about 5 weeks after customer approval of HR vector design.
Even though gene knock-outs can result from double strand breaks (DSBs) caused by Cas9 alone, SBI recommends the use of HR donor plasmids for more efficient and precise mutation. HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events. In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5´ or 3´ homology arms, can direct specific, targeted gene edits.
See link below for further information about SBI´s HR donor vector selection.
Custom CRISPR Cell Line Engineering
Please check with us if your cell line of choice is in stock at SBI enabling them to start your project right away at the prices shown below.
When it is not in stock, you can either provide it to us and we will ship it to SBI at a nominal shipping charge, or SBI can purchase it locally and we will charge you their purchasing costs.
SBI will use custom gRNA Cas9 and HR donor constructs to engineer target cell lines for knock-out, knock-in, tagging, or single nucleotide modification applications. Use of HR Donor is required, and is typically ordered as a package with the gRNA construct.
SBI can screen resistant cells to identify a clonal line with the desired modification (Cat# CS715B-1-SBI) or deliver the unscreened mixed cell population (Cat# CS715A-1-SBI).
For further details see individual service description.
Effective, efficient knock-out of miR-21 in HEK293 cells.4 gRNA, HR Donor design (with puro and GFP selection markers), implementation, and analysis performed by SBI’s genome engineering services team. (A) Low relative levels of miR-21—as measured by qPCR in GFP-positive clones—demonstrate the effectiveness of the approach. (B) After excision with Cre recombinase, the inserted GFP marker is efficiently excised, leaving only a single LoxP site from the HR Donor.
Source Data: Ho, TT, et al. Targeting non-coding RNAs with the CRISPR/Cas9 system in human cell lines. Nucleic Acids Res. 2015 Feb 18; 43(3):e17.
|Custom gRNA Design and Cloning||CS700A-1-GVO-SBI||1 ug DNA and bacterial streak||please inquire €||DETAILS||Add to Cart|
|Custom HR Donor Design and Cloning||CS600HR-1-GVO-SBI||1 ug DNA and bacterial streak||please inquire €||DETAILS||Add to Cart|
|Custom CRISPR Cell Line Engineering - Mixed Population||CS715A-1-GVO-SBI||1 x 10^6 cells||please inquire €||DETAILS||Add to Cart|
|Custom CRISPR Cell Line Engineering - Clonal||CS715B-1-GVO-SBI||1 clone||please inquire €||DETAILS||Add to Cart|