Easi-CRISPR Gene Editing

Easi-CRISPR is an improved gene editing technology that supports the specific insertion of large segments of exogenous DNA, generating knock-in mice for some applications much more efficiently than CRISPR or standard gene targeting methods.

  • Short Tag Knock-ins
  • Cre Knock-in Mice
  • Reporter Gene Knock-ins
  • Luciferase Reporting
  • Antigenecity-supporting sequences

Performance/Cost Ratio Excellent     Good    OK   Poor
  CRISPR/Cas9 Easi-CRISPR ES Targeting
Constitutive Knockout        
Point Mutation Knock-in         
Short Tag Knock-in         
Cre Recombinase
Knock-in
      
Reporter Knock-in       
cDNA Knock-in      
Conditional Knockout      
Genomic Replacement    
Applied properly, Easi-CRISPR is a robust and comparatively inexpensive method for exogenous DNA insertion in preclinical animal models.

Contact Taconic Biosciences about applying Easi-CRISPR technology to your animal model design program.

Easi-CRISPR Applications

Easi-CRISPR has been proven effective and efficient for the following genetic modifications:

Easi-Cre

The expression of a recombinase is used to delete a genomic region flanked by specific recognition sites (i.e. LoxP sites for the Cre recombinase). The sequence coding for the Cre recombinase is inserted in a specific genomic locus to achieve tissue-specific expression. The Cre cDNA can be inserted either at the 5' or the 3' of the gene (as shown in the example below).

Generating Cre knock-in mice with Easi-CRISPR A self-cleaving 2A peptide coding sequence is usually inserted between the Cre sequence and the target gene sequence, to ensure proper expression of the recombinase.

Easi-Transgene

Transgenic mouse models express exogenous DNA sequences inserted into their genome. For targeted transgenesis, an expression vector -- a human gene in this example) is inserted into a mouse genomic region lacking functional genes in order to avoid disruption of any endogenous function.

Generating transgenic mice with Easi-CRISPR

Easi-Flp

The expression of a recombinase is used to delete a genomic region flanked by specific recognition sites (i.e. Frt sites for the Flp recombinase). The sequence coding for the Flp recombinase is inserted in a specific genomic locus to achieve tissue-specific expression. The Flp cDNA can be inserted either at the 5' or the 3' of the gene (as shown in the example below).

Generating Flp recombinase mice with Easi-CRISPR A self-cleaving 2A peptide coding sequence is usually inserted between the Flp sequence and the target gene sequence, to ensure proper expression of the recombinase.

Easi-EGFP

Reporter genes are used to identify cells expressing specific genes. The sequence coding for the EGFP reporter protein, a green fluorophore, is inserted in a specific genomic locus to achieve tissue-specific expression. The EGFP cDNA can be inserted either at the 5' or the 3' of the gene (as shown in the example below).

Generating EGFP reporter mice with Easi-CRISPR A self-cleaving 2A peptide coding sequence is usually inserted between the EGFP sequence and the target gene sequence, unless it is desirable to express a fusion protein for intracellular localization studies.

Easi-mKate2

Reporter genes are used to identify cells expressing specific genes. The sequence coding for the mKate2 reporter protein, a far-red fluorophore, is inserted in a specific genomic locus to achieve tissue-specific expression. The mKate2 cDNA can be inserted either at the 5' or the 3' of the gene (as shown in the example below).

Generating mKate2 reporter mice with Easi-CRISPR A self-cleaving 2A peptide coding sequence is usually inserted between the mKate2 sequence and the target gene sequence, unless it is desirable to express a fusion protein for intracellular localization studies.

Easi-Luciferase

Reporter genes are used to identify cells expressing specific genes. The sequence coding for the luciferase reporter protein (Fluc), a light-emitting enzyme, is inserted in a specific genomic locus to achieve tissue-specific expression. The luciferase cDNA can be inserted either at the 5' or the 3' of the gene (as shown in the example below).

Generating luciferase reporter mice with Easi-CRISPR A self-cleaving 2A peptide coding sequence is usually inserted between the luciferase sequence and the target gene sequence to ensure proper expression of the reporter protein.

Easi-Tag

Proteins are tagged with short aminoacidic sequences to increase their antigenicity in antibody-dependent assays (e.g., Western blot, ELISA, Immunohistochemistry, Immunofluorescence, FACS, etc.). The sequence coding for the tag (usually HA, Flag™, or V5), is inserted in a specific genomic locus adding the desired aminoacids to the endogenous protein sequence.

Tagging proteins for antigenicity in knock-in mice with Easi-CRISPR The tag sequence can be inserted either at the 5' or the 3' of the gene.

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