Genome Editing in Drug Discovery. Группа авторов
4.2 Genome editing service providers.
| Company | Location | Technologies | Services |
|---|---|---|---|
| Applied Stem Cell | Milpitas, CA | CRISPR/Cas9 | Cell engineering: cell lines, ES/iPSCs |
| ATCC | Manassas, VA | CRISPR/Cas9 | Cell engineering: cell lines |
| BioGene | Shirley, NY | CRISPR/Cas9 | Cell engineering: cell lines, iPSCs, primary cells; rodent models; plants models |
| Canopy Biosciences | St. Louis, MO | CRISPR/Cas9 | Cell engineering: cell lines, ES/iPSCs |
| Cellecta | Mountain View, CA | CRISPR/Cas9 | genetic screening |
| Cellular Dynamics International | Madison Wisconsin | CRISPR/Cas9 | ES/iPSC engineering |
| Charles River Laboratories | Wilmington, MA | CRISPR/Cas9 | Cell engineering; rodent models |
| Crown Bioscience | Santa Clara, CA | CRISPR/Cas9 | Rodent models |
| DefiniGEN | Cambridge, UK | CRISPR/Cas9 | iPSC engineering |
| Evotec | Hamburg, Germany | CRISPR/Cas9 | In vitro genetic screening |
| genOway | Lyon, France | CRISPR/Cas9 | Cell engineering: cell lines, rodent models |
| GenScript | Jiangning, China | CRISPR/Cas9 | Bacterial engineering; cell line engineering |
| Horizon Discovery | Cambridge, United Kingdom | CRISPR/Cas9 | Cell engineering: cell lines, ES/iPSCs; in vitro and in vivo genetic screening |
| Merck KGaA | Darmstadt, Germany | CRISPR/Cas9, ZFN | Cell engineering: cell lines, iPSCs, primary cells |
| Oxgene | Oxford, United Kingdom | CRISPR/Cas9 | Cell line engineering |
| Synthego | Redwood City, CA | CRISPR/Cas9 | Cell engineering: cell lines, iPSCs, primary cells; in vitro genetic screening |
| System Biosciences | Palo Alto, CA | CRISPR/Cas9 | Cell line engineering |
| ThermoFisher | Carlsbad, CA | CRISPR/Cas9, TALEN | Cell line engineering |
| Ubigene | Guangzhou Science City, China | CRISPR/Cas9 | Bacterial engineering, fungal engineering, cell line engineering |
| WuXi AppTec | Shanghai, China | CRISPR/Cas9 | Rodent models; in vitro genetic screening |
4.3.3 Next‐Generation in vivo CRISPR Screening
The CRISPR toolkit for genome editing has been rapidly evolving and next‐generation CRISPR technologies have been applied for genetic screening in vivo. For example, in vivo screens based on CRISPRa (Braun et al. 2016; Ebright et al. 2020), CRISPRi (Li et al. 2020), exon excision (Thomas et al. 2020), and Cas12a/Cpf1‐mediated double gene knockout (Chow et al. 2019) have been reported. Most recently, pooled in vivo CRISPR screening has been combined with single‐cell RNA‐seq analysis so that the relationships between genotypes to in vivo phenotypes can be mapped at single‐cell resolution (Giladi et al. 2018; Jaitin et al. 2016). We anticipate this approach to be coupled with diverse functional readouts across a broad spectrum of animal models and continue to transform biomedical research and drug discovery.
4.4 Working with Service Providers for Outsourcing CRISPR Studies
This section covers CRISPR outsourcing solutions, but the same principles apply to other forms of gene editing technologies like TALENs and ZFNs. Although the tools and methods for application of genome editing technologies are becoming more accessible, some labs may choose to outsource partial or entire experiments on a fee‐for‐service basis. Genome editing CROs have invested considerable efforts on internal development and have amassed great knowledge on how the technology works, which translates to a high competence to design genome editing projects and troubleshoot problems. In addition, CROs enable researchers to conduct research without the need to purchase and maintain expensive equipment (e.g. flow sorter, or next‐generation sequencing instruments). CROs are also used when there is time pressure to deliver while internal resource is in short supply, or in cases where there is limited or no internal expertise. Such a business model has several advantages and disadvantages which need to be considered before proceeding with this option (Pichler and Turner 2007).
4.4.1 Critical Steps for Outsourcing
A number of companies and academic institutions offer genome editing services. Providers can be found through websites including Science Exchange (https://www.scienceexchange.com) and Biocompare (https://www.biocompare.com) and we provide a non‐exhaustive list in Table 4.2. It is important to base your choice on the reputation and track record of the company, otherwise you run the risk of contracting a CRO that will use your project for its own capability development (Pichler and Turner 2007). If possible, it is advisable to visit the CRO facility to inspect their laboratory set‐up, view the equipment used, and discuss case studies that the company has completed. Also, you will need to perform due diligence checks. For example, discuss if the CRO holds foundational CRISPR licenses and understand their workflows with regard to data integrity (i.e. how they record and track their data). The next step is to set up a Confidential Disclosure Agreement (CDA) so that you are able to freely exchange information about the project you want to conduct and codevelop a detailed experimental plan. As in the case of sourcing reagents, when discussing