Knockout Mouse Catalog | Cyagen APAC

CRISPR/Cas Knockin Mice

CRISPR/Cas9 Knockin Mouse Services
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Cyagen Biosciences offers CRISPR-based, large-fragment knockin, at any locus, and conditional knockin mice generation in as little as 4 months, at highly competitive prices. We guarantee delivery of a minimum of 2 founders or 3 F1 animals for knockin fragments up to 4 kb (up to 3 kb for conditional knockin projects). Projects with larger knockin/cKO fragment will be evaluated case by case. Tell us the name of gene of your interest (for knockin project, target site you wish to target), and we will generate your mouse model.


◆ Workflow of CRISPR/Cas Knockin Mice (Any Locus)/Conditional Knockin Mice

CRISPR/cas9 knockin mice (any locus)

◆ Description of Services

Any-locus large-fragment knockin/Conditional knockin strategy design

Tell us the name of gene of your interest and we will design a nuclease-mediated strategy for you. This includes design of the donor vector and gRNA vectors, and selection of target sites in the gene based on our optimized algorithm that maximizes on-target nuclease activity and minimizes off-target activity. Genotyping assays based on PCR and sequencing will also be designed for the screening of founder mice.


CRISPR Knockin/Conditional knockin vector construction

DNA vectors that express the desired nucleases and donor vector will be constructed. Where needed, the efficacy of these vectors will be tested in cell culture.


CRISPR/Cas injection into mouse eggs

mRNA preparation: Nuclease expression vectors will be transcribed in vitro. The resulting mRNA will be artificially capped and polyadenylated to facilitate proper translation in mammalian cells.


Nuclease injection to obtain founders: The nuclease mRNA and donor vector will be co-injected into fertilized mouse eggs, followed by implantation of the eggs into surrogate mothers to obtain offspring. In cases where the nuclease expression vectors are designed and constructed by Cyagen, we will inject as many eggs and/or target as many sites as needed to fulfill the guarantee. In cases where the nuclease expression vectors (or their mRNA products) are provided by the customer, we will inject a minimum of 200/300 eggs (based on strain) and screen pups for founders carrying desired mutation. If no founders are identified, more injections can be performed at an additional charge.


Founder screening

Pups will be screened by PCR to identify knockin/floxed founder mice. Specifically, the site targeted by CRISPR will be PCR-amplified to reveal any insertions. Occasionally, an animal may be found to have both alleles of the target site mutated. For conditional knockin projects, mice carrying two loxP sites on the same allele flanking critical exon(s) are considered floxed mice.


Breeding founders to obtain F1

For some projects, the generation of founder mice is the end point. However, some customers wish to have us breed the founders further to obtain F1 mice. Cyagen will breed up to 3 founders to wildtype mice of matching strain background, and genotype their offspring to obtain F1 mice bearing the large-fragment knockin/floxed allele.


◆ Guarantee

Cyagen offers the best guarantee in the industry – we will fully refund the client’s service fee if animals with the specified genotype are not generated (except for genetic modifications severely affecting viability, morbidity, or fertility). Given the complexity of biological systems, a particular genetic modification may not result in the desired phenotype. As such, Cyagen's guarantee covers the creation of animals with the specified genotype, not a particular phenotypic outcome in terms of transcription, protein/RNA function, or organismal biology.

Why work with Cyagen?

  • 14+ years' experience in custom animal model generation
  • 78,000+ animal models generated and delivered worldwide
  • 4,700+ citations in SCI journals for our services
  • 3,000+ university and company partners
  • 100% money-back guarantee option available for most services
  • AAALAC-Accredited and OLAW Assured
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