Package Option
The MGEF will perform all the steps necessary to generate your CRISPR mouse model. This includes:
- Allele design
- CRISPR reagent procurement and testing
- Microinjection
- Founder screening
- Breeding and sequence analysis of N1 generation heterozygous mice
- Off-target analysis
- Development of an automated genotyping strategy in conjunction with Transnetyx
Please contact Bill Shawlot to schedule a meeting to discuss the specifics of your project.
Microinjection Option
Investigators can also submit CRISPR reagents of their own design for the core to microinject.
- Check with Bill Shawlot for CRISPR design considerations. We use the CRISPOR gRNA design program.
- The MGEF recommends purchasing gRNAs from IDT or Synthego.
- We suggest purchasing ALT-R HDR Donor oligos from IDT for introducing point mutations or short insertions. For larger inserts, Alt-R HDR Donor Blocks are recommended.
- Labs should submit the lyophilized gRNAs and oligos to the MGEF. The core will resuspend the reagents and mix the components at the appropriate concentration for microinjection or electroporation.
Expectations
- The core will guarantee to inject a specific number of embryos based on the anticipated efficiency of the editing event. Because the efficiency of Homology Directed Repair editing events can be variable, additional rounds of microinjection or electroporation may sometimes be necessary. The Director will meet with the lab to discuss options in these instances.
- If the gene-editing event appears to cause embryonic lethality, the core will meet with the investigator to review the microinjection data and discuss alternatives.
- For the model package option, the core will bill the investigator after completing each service step.