E-Book 2nd Congress

  • Application of the CRISPR/Cas9 System in transgenic animals
  • sadaf safaie,1,* Hamid Mir Mohammad Sadeghi,2 sadaf safaie,6
    1. Department of pharmaceutical biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran
    2. Department of pharmaceutical biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran
    6. Department of pharmaceutical biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran


  • Introduction: In recent years, novel genome editing technologies based on the CRISPR-associated RNA-guided endonuclease Cas9 has provided the ability to rapidly and economically introduce sequence-specific modifications depends on the generation of double-strand break (DSB) and DNA repair process into the genomes of cell and organisms. Gene modification can be introduced into the animal genome through homologous recombination and embryonic stem cell technology. Genetically modified animals, especially gene knockout editing is one of great interest in the prevention and treatment of human diseases. In this review, we discuss the CRISPR/Cas9-based transgenic models to enhance engineering a wide spectrum of mutations found in human cancers.
  • Methods: CRISPR/Cas system is the most flexible and user-friendly platform to generate transgenic animals. Non-viral vectors, viral vectors, and physical delivery are the most widely used method for delivery of CRISPR/Cas9 for genome editing. The physical delivery, including microinjection, electroporation and somatic cell nuclear transfer (SCNT) procedure are widely used for target delivery of CRISPR/Cas system to generate transgenic animals.
  • Results: CRISPR/Cas9 technology has been used to generate target genome modifications. Gene modification can be introduced into the animal genome through homologous recombination and embryonic stem cell technology. The discovery of site-specific endonucleases has provided the ability to establish deletion (Knock Out) or insertion (Knock In) of specific genomic sequences on a single step, directly applied to zygotes. Genetically modified animals, especially gene knockout editing is one of great interest in the prevention and treatment of human diseases. Methodology delivery of crispr/cas system: Viral vectors are efficient in gene delivery, but they have some contraindication due to many drawbacks such as off-target effect, immunogenic and inflammatory responses, limited packaging capacity, and high cost in production. Non-viral vectors, including Nano carriers and nanoparticles such as Nano polymeric- and lipid-based structures, rigid nanoparticles, nanoparticles coupled to specific ligand systems including arginine–glycine–aspartate (RGD) peptide, porous silicon, mesoporous silica, metal–organic, cell-penetrating peptides. This method is efficient but expensive. The physical delivery, including microinjection, electroporation and hydrodynamic delivery show high efficiency for the application in vitro, but not satisfy for in vivo application especially in large animals.
  • Conclusion: The simplest and the most flexible and user-friendly platform to engineering nuclease system to generate transgenic animals is the CRISPR/Cas9 system. Genetically modified animals, especially gene knockout models, have been valuable for mimic human disease. This system provides multiplex genome engineering, based on site-specific recombinases, primarily the Cre-LoxP system, enables the creation of animal transgenic models, have the capacity to engineer a wide spectrum of mutations found in human cancers.
  • Keywords: Genome Editing tools, CRISPR/Cas9, Transgenic animals