Applying CRISPR Cas9 Technologies

The relative ease of genetic engineering with CRISPR is transforming the development of more complex disease-relevant cell-based assays to improve predictability for drug therapies. CRISPR Cas9 design tools can be applied to multiple phases of the drug development process. Gene knock-outs and gene knock-ins introduced in cell-based screening assays create a system that best mimics a disease state. The ability to turn on and off genes using CRISPR Cas9 has been used to distinguish between on-target and off-target effects in target validation assays.

Value of CRISPR Cas9 Technology in Phenotypic Rescue Experiments

Added value of CRISPR Cas9 technology in phenotypic rescue experiments

Fail early, fail fast using a phenotypic approach to target identification and hit validation. Read the Article

CRISPR Cas9 in High-Throughput Screening

Through CRISPR Cas9 engineering, a multitude of high-throughput screens (HTS) can be designed to mimic the most specific of targets in disease-relevant cells. Charles River employs CRISPR technology to develop both pooled and arrayed HTS. Pooled screening generates relatively clear-cut readouts, such as cell proliferation, cell death or sortable marker proteins. For more complex phenotypic readouts, arrayed screens are developed incorporating high content imaging. Arrayed CRISPR libraries are now emerging with the guide RNA being either expressed from lentivirus or produced as a synthetic molecule. The specificity of these screens results in stronger hits.

High Content Screening CRISPR Cas9 Generated Cells

Disease relevant in vitro assays for ALS in motor neurons derived from control patient iPSCs.

Disease-relevant in vitro assays for amyotrophic lateral sclerosis in motor neurons derived from control and patient iPSCs. Download the Poster

CRISPR Cas9 for Target Validation

The power of the CRISPR technology is its ability to precisely create the target of interest when developing assays. In target validation, the CRISPR technology has the advantage over RNAi to fully abolish gene expression as opposed to the transient and incomplete silencing of the target as achieved by siRNAs. This also has a profound impact on rescuing the phenotype to confirm target specificity by adding back methodology, which will give far better interpretable results in case of a complete knockout by CRISPR as opposed to an incomplete silencing by RNAi. Charles River has developed a CRISPR phenotypic rescue assay to validate the interaction between a compound and the intended drug target to discriminate between on-target and off-target effects. Validating your compound using CRISPR Cas 9 technology produces stronger lead candidates.

Validating Targets Using CRIPSR/Cas9 Repair

Disease relevant in vitro assays for ALS in motor neurons derived from control patient iPSCs.

Validation of the interaction between a candidate compound and the intended drug target by a phenotypic rescue approach. Download the poster

Additional CRISPR Resources

CRISPR is revolutionizing cell-line development. Have questions, or simply want a better understanding of CRISPR technology? Check out this helpful resource from our friends at The Broad Institute. Visit their pages for a brief history, research highlights, and information on notable CRISPR scientists.

Need a CRISPR mouse or rat model? Check out our diverse collection of Research Models or send us your specifications.


Frequently Asked Questions (FAQs) for CRISPR Cas9

  • What is the main function of the CRISPR Cas9 system?

    CRISPR technology exploits the Cas 9 gene that enables cells to respond to and eliminate invading genetic material. Using Cas9, gene sequences can be knocked-in or knocked-out creating unlimited potential for disease-relevant cell lines for use in drug discovery and gene therapy.

  • What is the benefit of using CRISPR Cas9 over other genome editing technologies?

    The beauty of the CRISPR Cas9 system is that an RNA molecule is driving the specificity of the system and these molecules are easy to design and to produce. Targeting the mRNA to change the properties of a cell is a powerful tool since the host genome is unaffected.

  • Are there CRISPR technologies besides Cas9?

    New Cas9-like proteins are still being discovered with different properties and requirements, thereby expanding the CRISPR toolbox for, what seems like, unlimited possibilities in genome editing and gene regulation. Cas14 is one such example. Isolated from archaebacteria, it is smaller than Cas9 and can also cut ssDNA. Another Cas9 variant was reported with minimal PAM requirements, meaning more parts of the genome can be targeted.

+ CRISPR-Cas9 used under licenses to granted and pending US and international patents from The Broad Institute and ERS Genomics Limited.

Consult a CRISPR Technology Specialist