Explore UAB


{slide=Complete CRISPR model design and creation}

  • Guide design and preparation
  • Blastocyst assay to assess activity of guides
  • Establishment of genotyping strategy
  • Repair template design
  • Compatible with most efficient guide(s)
  • Unique restriction site for downstream screening of founders
  • Pronuclear microinjection of 150 embryos
  • Transfer of embryos to pseudopregnant recipients
  • Optional screening of founder animals
  • Transfer of founder litters at weaning




Rodent Model Generation

{slide=Pronuclear Microinjection}


  • Loss of function alleles
  • Knockout
  • Large genomic deletions
  • Hypomorphic

Targeted Insertions

  • Patient mutations
  • SNPs
  • Conditional alleles
  • ROSA26 insertions
  • Reporter tags

Simple Transgenics

  • Reporters
  • Overexpression vectors


{slide=ES Cell Microinjection}

ES Cell Targeting

  • Conventional
  • CRISPR induced recombination
  • CRISPR guides and compatible ROSA26 targeting vector


Preparation of commercially available ES cells (EUCOMM / KOMP)

Assisted Reproductive Techniques



  • Sperm
  • Embryos
  • Oocytes
  • Ovaries


  • Sperm
  • Embryos




  • In vitro fertilization
  • Transfer of cryopreserved embryos
  • Pathogen-free
  • Ovarian transplant


  • Transfer of cryo-preserved embryos



  • Design of transgenic constructs
  • Screening strategies
  • Breeding schemes

Maintenance of Stock Lines

  • Cre stains (ex. Ella)
  • Reporters (ex TdTomato, GFP)
  • Male/female breeders and controls (ex. C57B6J)