Services Provided by the Genome Editing Core

The GESR offers the following services (more to follow):

CRISPR/CAS9 -  Mediated Gene Editing in Mouse

The CRISPR-Cas9 system has revolutionized the production of genetically modified rodent models.  Following are examples of approaches we can take to provide nearly any type of experimental system:

  • Gene Knock-out: Straight knock-outs of genes can be accomplished simply by electroporating mouse zygotes with CRISPR guide RNAs complexed with Cas9 protein causing a site-specific DNA double strand break, generally resulting in small deletions and disruption of the coding sequence thus rendering proteins defective.
  • Gene Replacement (amino acid changes): Specific amino acid changes to coding sequences and SNP (single nucleotide polymorphism) modeling is done using a single stranded DNA oligonucleotide bearing the change(s) along with the CRISPR-Cas9 complex. This approach will greatly assist translational research using models based on human disease variants.
  • Conditional Models: Conditional mutants facilitate studies of tissue-specific gene ablation and can be made using single stranded DNA oligonucleotide donors containing loxP sites which direct Cre-mediated gene deletion.
  • Epitope tagging, including fluorescent protein tagging: Fluorescent/epitope tags and expressing cassettes (Cre, rtTA, Flp) can be inserted into endogenous genetic loci to facilitate identification of proteins in vivo or direct expression of in vivo modifying tools in a tightly regulated manner.

DNA Microinjection for transgenic generation

  • Transgene DNA Purification available as an additional service

BAC DNA Microinjection for BAC transgenesis

ES Cell Line Injection

  • ES Cell preparation and Karyotyping available as additional services

CRISPR Target Validation (for Gene Editing in Mouse)

Mouse Cryopreservation:

  • Sperm Cryopreservation
  • Embryo Cryopreservation

In Vitro Fertilization

  • Frozen/Fresh sperm for line import, recovery or rapid expansion

Mouse Line Rederivation

  • Live Mice as Source
  • Frozen Embryos as Source

Genotyping

  • The GESR offers genotyping of lines generated as an additional service.
  • The GESR will design and validate genotyping assays as needed.

 

Fees for GESR services are available upon request. Please email: gecf-ru@rutgers.edu