CAS9 Delivery  

CAS9 Delivery refers to the application of affinity-functionalized nanotechnology to deliver CRISPR-associated protein 9 (Cas9) complexes for genome editing. This method uses engineered nanoparticles conjugated with specific affinity ligands to target and deliver Cas9 ribonucleoprotein (RNP) complexes or nucleic acids encoding Cas9 into designated cells or tissues with high precision. 

Technical Content and Mechanism

• Nanoparticle Design for Cas9 Delivery:
  • Material Selection: Nanocarriers such as lipid nanoparticles (LNPs), polymeric nanoparticles (e.g., PLGA, polyethyleneimine-based), or inorganic nanoparticles (e.g., gold or silica-based) are chosen based on biocompatibility, cargo capacity, and ease of surface modification.
• Affinity Functionalization for Targeting:
  • Ligand Conjugation: The nanoparticle surface is modified with affinity ligands (antibodies, peptides, or aptamers) that specifically bind to surface markers on target cell populations. Techniques such as carbodiimide chemistry (EDC/NHS coupling), thiol–maleimide reactions, or click chemistry are used for stable ligand attachment.
• Cas9 Complex Loading and Protection:
  • Cargo Encapsulation or Conjugation: Cas9 RNP complexes, plasmids encoding Cas9, or mRNA for Cas9 are loaded into the nanoparticle core or attached to the surface. Encapsulation protects Cas9 from degradation and shields it from the immune system.
• Gene Therapy:
  Targeted delivery of Cas9 RNPs using AffiNANOTECH can correct disease-causing mutations in genetic disorders such as muscular dystrophy, cystic fibrosis, or hemophilia by editing patient-specific genes in affected tissues.
• Cancer Research and Therapy:
  Using nanoparticles to deliver Cas9 complexes specifically to tumor cells can enable targeted gene disruption of oncogenes or restoration of tumor suppressor functions, reducing systemic toxicity and improving therapeutic outcomes.
• Neurological Disorders:
  By designing nanoparticles to cross the blood–brain barrier and target neuronal cells, AffiNANOTECH platforms can deliver CRISPR components for editing genes related to neurological conditions like Huntington’s disease or Alzheimer’s disease.

CAS9 Delivery AffiNANOTECH leverages the precision of affinity-based nanoparticle targeting to enhance the delivery of CRISPR/Cas9 systems for genome editing. Through careful nanoparticle design, surface functionalization with specific affinity ligands, and controlled release mechanisms, this approach aims to achieve efficient, cell-specific gene editing with reduced off-target effects and improved safety profiles, opening new avenues in gene therapy, precision medicine, and biomedical research.