Custom Mouse Injection-Ready Kit (Validated)

Injection-Ready Validated CRISPR Kits for Mouse Model Generation

CRISPRCRITTERS Injection-Ready Kit is a CRISPR kit for the generation of transgenic mice and rats. This CRISPR mouse model kit contains Surveyor Assay validated gRNAs designed to target your gene of interest, donor DNAs and microinjection-validated Cas9 mRNA. For customers who prefer a more stringent off-target profile, we also provide mRNAs of Cas9 D10A or H840A nickase.

  • Injection-Ready
  • Validated in Surveyor Assay
  • Cas9 Nickase Available
  • Full Mouse Generation Service Available

Cas9 Variants

Rat Model Generation

CRISPR Kit

 

 

An Example of gRNA Validation Data (mRosa26 locus)

Two gRNA sequences were cloned into gRNA-expressing vectors individually. Each construct was transfected into mouse N2A cells, individually. gRNA activity was determined by SURVEYOR assay on the PCR products, using the SURVEYOR Mutation Detection Kit, according to the manufacturer’s instructions. The results showed indel frequency of 26% and 12% for the two gRNA constructs, respectively.

An Example of Generating a Point Mutation Mouse Model Using Cas9 D10A nickase CRISPRCRITTERS™ Mouse Point mutation kit

The mixture of two gRNAs, a single-stranded DNA oligo containing desired mutation, and Cas9 D10A nickase mRNA was injected into cytoplasm of mouse embryos. PCR products from genomic DNA were sequenced. Results from a positive pup with desired T>A mutation are shown below.

 

An Example of Generating a Conditional Knock-out Mouse Model Using Cas9 CRISPRCRITTERS™ Knock-in kit

A mixture of active guide RNA (gRNA), a single strand oligo donor containing a HA tag sequence and Cas-9 mRNA was injected into the cytoplasm of C57BL/6 embryos. Mice born from the microinjected embryos were screened using PCR-restriction digest analysis. Pups #2, 6, 13 and 17 showed heterozygous insertion. PCR products were topo cloned and sequenced to confirm the HA tag was inserted at right location.  Positive animals were further verified by sequencing the region adjacent to the targeted location.

 
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