More from the RapidGen™ Platform
Figure 1. FastGenTM Model of Macrophage Knockout in Zebrafish. Knockout of the irf-8 gene that encodes Interferon Regulating Factor 8 which promotes macrophage development.
(A) Uninjected sibling control Zebrafish embryos at 48 hours post injection with macrophages illuminated by expression of mfap4:GFP.
(B) FastGenTM Model injected with irf-8 Cas9 and multiple sgRNAs showing visible reduction of GFP+ macrophage cells. Co-injected with Rhodamine dye to indicate successful injection.
InVivo Biosystems optimized the crispant approach to efficiently “break” a gene at multiple sites using multiple sgRNAs, particularly in the context of zebrafish knockout studies. The result is a highly penetrant gene knockout that can be assayed by phenotype immediately in the F0 animals (Figure 1). A traditional CRISPR-based edit modifies a single site in a gene and requires months of propagation and screening to stabilize and confirm the edit before assaying. Unlike transient knockdowns, such as morpholinos, the FastGen™ modifications are permanent and can be propagated to establish a stable line.
Deliverables
Because the FastGenTM approach creates a transient model, phenotypic analysis is performed by InVivo Biosystems and animals are not delivered to the customer. Based on the phenotypic screen chosen, deliverables might include:
Determine whether a compound or formulation induces liver damage. Measurements using early life-stage zebrafish include gross tissue health, necrosis, liver function and cell death.
Assess whether a compound or formulation induces nervous system damage. Early life-stage zebrafish will be used to measure neural gene expression via qPCR.
Cardiotoxicity Testing
Assess whether a compound or formulation induces heart damage in zebrafish. Early life-stage zebrafish will be used to assess cardiac function.
Introduction
We will set up a 30-minute call with you to understand your needs. During this call, we will define the scope of your project, answer your practical questions and help you assess whether our service is a good fit for you.
Design
After we decide on the scope of the project, we will create a proposal that includes cost and time estimate for each experiment proposed. You will be able to personalize your project plan.
Logistics
We will send you a final statement of work and payment schedule. Once we receive your first PO, we will start the experiments and give you a defined timeline for your project.
Report Out
We will keep you updated every two weeks on the status of your project. At the end of your project, we provide you with a report that includes comprehensive findings and key takeaways.
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Built on a robust portfolio of CRISPR genome editing patents, InVivo Biosystems has established a comprehensive preclinical discovery and development platform. By leveraging alternative models and proprietary methods, we accelerate go-to-market timelines for pharmaceutical companies and beyond.