Large Insertions

We are now producing genome modified mice using a groundbreaking technology published as “CRISPR-READI.” This method (PMID:31242412) combines viral delivery of donor DNA into embryos with electroporation of CRISPR/Cas9 ribonucleoproteins. The results have been nothing short of remarkable.

• Efficiency: CRISPR-READI boasts exceptional efficiency. In a single delivery attempt, we’ve successfully attempted and completed fourteen mouse genome modification projects, resulting in an average of 42% of pups having the desired allele!
• Expanded Possibilities: With AAV’s upper packaging limits of approximately 4.7 kb, we can now create mice with floxed alleles or that express degrons, recombinases and fluorescent proteins.
• Longer Insertions: Efficiencies are high enough that we can produce longer insertions by introducing two or more viruses containing distinct, overlapping DNA donors.

Workflow:

Advisory Meeting → Strategy Design → Reagent Procurement → Editing of Mouse Embryos → Screening of F0 Generation → Breeding and Screening of N1 Generation

Average Time to F0 Founders:

5-6 months

Price through F0 Founder Generation:

Shared resources must adjust pricing annually. Projects are by custom quote. Most full-service projects we offer fall within a range of $20,000-25,000.

Success Rate:

100% for edits that do not severely affect viability, morbidity, or fertility

Genotype Guarantee:

As a shared resource, we cannot offer a money back guarantee. Based on previous projects, we can confidently state that large knock-ins that do not severely affect viability, morbidity, or fertility, that are introduced into the standard strains, C57BL/6J or C57BL/6N, will be produced within two electroporation attempts, and most within a single attempt.

Models Produced: 14