The latest tool in genome editing – CRISPR/Cas9 – allows for specific genome disruption and replacement in a flexible and simple system resulting in high specificity and low cell toxicity. The CRISPR/Cas9 genome editing system requires the co-expression of a Cas9 protein with a guide RNA. With the protospacer-adjacent motif (PAM – the sequence NGG) present at the 3′ end, Cas9 will unwind the DNA duplex and cleave both strands upon recognition of a target sequence by the guide RNA. The functional cassette in the rescue donor vector can then be inserted into the unwound DNA. The repaired genome will now express your desired sequence with or without tags.
OriGene provides all the vectors and service for the Cas9-genome editing. http://www.origene.com/CAS9/
pCas-Guide vector (SKU# GE100002) encodes Cas9 enzyme and the guide RNA. You can use OriGene free gRNA design tool to design your guide RNA. Have them synthesized by any commercial company such as IDT . Anneal them together to make a double strand DNA and ligate the dsDNA to pCas-Guide vector. OriGene also offers an annealing buffer (SKU# GE100007) for you to make the dsDNA.
pCas-Scramble (SKU#GE100003) contains a scrambled sequence in pCas-Guide vector. It is a good negative control that you can use for your work.
The gene specific rescue donor vector can be made through gene synthesis (www.blueheronbio.com) or from your own donor constructs. OriGene also provides donor vector construction service. To order your donor vector service, please go to http://www.blueheronbio.com/Services/Genome-Editing.aspx
You can also select the following premade functional cassettes:
Cas9 Further Reading
DreamFect™ Stem (TT100159 & TT100160) is an original, simple and efficient method especially developed to transfect multipotent stem cells with high transfection efficiency and superior transgene expression level.
1. High transfection efficiency for multipotent Stem Cells
2. Minimized toxicity due to reagent biodegradability and low DNA amount required
3. Do not affect phenotype and differentiation potential
4. Serum Compatible
5. Simple, Ready-to-use and rapid (one vial, no specific buffer, no need magnetic plate)
Over 2,000 TrueORF Clones are available for viral protein expressions
- Covering over 120 human viruses
- Codon optimized for human cell expression
- All cloned in proven pCMV-Entry Vector
- Myc-DDK tagged for convenient detection and purification
- 60 destination vectors for additional applications
OriGene now offers the service to generate Mass Spectrometry (MS) standards for 6,000 human proteins. Produced in human HEK293T cells and labeled with [U- 13C6, 15N4]-L-Arginine and [U- 13C6, 15N2]-L-Lysine, the full length proteins with appropriate post-translational modifications are the best identification and quantification standards.
Supriority of OriGene’s MS standards:
- Spiking at the early stage of sample process for accurate quantification
- Identify the best SRM and MRM transitions through experimental data
Internal standards of quantitative MS in cancer biology
- Authentic posttranslational modifications by using human HEK293T cell line.
- Higher data consistency than synthetic peptide internal standard
- Suitable for all types of MS equipments
- Over 90% incorporation efficiency.
Are you sure that your IHC signals are truly specific?
High specificity is the pre-requisite for IHC antibody to be used for diagnostic and therapeutic applications. The cross-reactivity may potentially cause unexpected side effects and false diagnostic reports for clinicians. However the validation tools for antibody specificity are lacking. Although research data from various groups has shown that some monoclonal antibodies on the market are not mono-specific, it remains a challenge to produce the truly mono-specific antibodies to replace them.
OriGene is proud to announce that we have developed a high density protein microarray chip for antibody specificity testing. With the world’s largest collection of overexpression protein lysates, OriGene is in the unique position to produce a high density chip of human proteins. This protein chip is spotted with 10,464 (10K) unique over-expressed proteins in duplicate on a single nitrocellulose coated glass slide. This protein microarray technology has been used to validate the specificity of an existing ERCC1 diagnostic monoclonal antibody, and has been applied as a screening method to identify the most specific UltraMAB™ monoclonal antibody for this target.
The excision repair cross-complementation group 1 (ERCC1) protein is an important biomarker for clinicians to predict whether certain patient populations with non-small cell lung carcinoma will respond to cisplatin chemotherapy. As such, it is critical to develop highly-specific immunohistochemistry validated monoclonal antibodies for this diagnostic test. Several publications revealed that 8F1, the most commonly used antibody clone for ERCC1, exhibits cross-reactivity to an unknown protein in ERCC1 deficient cell lines. By using OriGene’s high-density protein microarray technology, the corresponding cross-reactive binding protein for 8F1 monoclonal antibody was identified. This technology also enable us successfully develop the most specific UltraMAB™ monoclonal antibody for ERCC1 (4F9). This data was further confirmed by western blot analysis.