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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

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