Customized CRISPR sgRNA and donor plasmids design and construction services
The type II CRISPR/Cas9 system of Cas9 from Streptococcus pyrogenes (SpCas9) can be programmed by a chimeric single-guide RNA (sgRNA), a hybrid duplex of crRNA (CRISPR RNA) and tracrRNA (trans-activating crRNA) to make precise double-stranded break (DSB) in the genome. The crRNA and tracrRNA can be fused together to create a chimeric, single-guide RNA (sgRNA), and further could be construct via a sgRNA-expression plasmid. On rare occasions, certain sgRNAs may not work for reasons yet unknown; therefore, we recommend designing at least two sgRNAs for each locus and test their efficiencies in the intended cell type.
CRISPR/Cas9 is very easy to customize as this system only requires two components, the endonuclease Cas9 and a guide RNA (gRNA).
Fusion Biolabs provides customized CRISPR constructs design and construction services
- sgRNA design and sgRNA-expression plasmid preparation
- Targeting donor plasmid design and preparation
I. Two Vectors based CRISPR/Cas9 genome engineering system
Fusion Biolabs has simplified the design and selection of the target sequence (gRNA) and clone into our pOK1-gRNA vector (Two-vector system) and we will provide you a complimentary Cas9 expression plasmid upon request.
II. One Vector based CRISPR/Cas9 genome engineering system
Fusion Biolabs has simplified the design and selection of the target sequence (gRNA) and clone into our pOK2-gRNA vector (One-vector system).
III. Donor plasmid construction with a predefined cassette
Site-specific genome editing tool, CRISP/Cas9 system can be customized to recognized and cleave a double-stranded DNA molecule (DSB) at a specific site, which can be subsequently repaired by endogenous cellular pathway of homology-directed repair (HDR). HDR can seamlessly repair DNA break in the presence of a DNA repair template with homologous regions spanning the cleavage site. The DNA template used in HDR can be customized by placing a GOI in between two homology arms (HAs) and then delivered exogenously in a donor plasmid.
The donor plasmid consists of the following fragments:
- plasmid backbone, which include elements for replication in the bacterial host;
- 5′ and 3′ homology arms (HA) ~750 bp each to mediate homology-directed targeted integration
- drug resistance cassette, harboring neomycin or other drug resistance gene together with the promoter and terminator sequences;
- gene expression cassette, harboring gene of interest (GOI) with the desired promoter and terminator sequences.
Ordering Information
Customized CRISPR sgRNA and Donor plasmid Design and Construction Services
Catalog | Description | Unit | Price |
---|---|---|---|
71101 | Target sequence gRNA cloning into pOK1-gRNA vector (2 vector system) | 1 Project | Contact Us |
71102 | Target sequence gRNA cloning into pOK2-gRNA vector (1 vector system) | 1 Project | Contact Us |
71103 | Donor vector design and construction with a predefined cassette | 1 Project | Contact Us |
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