CLICK GENE

SPLITTERA for Gene Therapy

Why SPLITTERA? CLICK GENE

SPLITERA is a versatile protein engineering tool that allows the conjugation of proteins through the formation of new peptide bonds. This technology address the main limiting factors in Gene Therapy. It overcomes gene size limitations of viral vectors, allowing the covalent reconstitution of large proteins expressed by different viral vectors in vivo. Moreover, it allows virus surface modification to target viral particles to specific tissues.

Key Advantages

  • UNIVERSAL:
    • SPLITTERA Split Inteins: protein elements that allow the reconstitution in vivo of large proteins expressed by different viral vectors.
    • Universal Platform: Standard conjugation reaction suitable for all antigens.
    • Overcome the challenge of protein size.
    • Minimalsequence requirement.

 

  • ADDITIONAL FEATURES:
    • Specificity: 100% specific.
    • Versatility: 4 different orthogonal pairs of inteins that can be co-expressed.
    • High reaction yield.
    • Extremely fast reaction.
    • Active under physiological conditions.

 

  • VALIDATED METHOD

Large gene delivery

SPLITTERA overcomes gene size limitations of viral vectors, allowing the covalent assembly of large proteins expressed by different viral vectors in vivo through Intein mediated trans-splicing reaction. Recent work demonstrate that inteins are applicable to genes related to inherited and non-inherited diseases1 and to deliver highly efficient base editors .

  • Co-infection of host cell with two (or more) independent viral vectors carrying each a fragment of the DNA coding for the POI (Protein of Interest) fused to the corresponding Splittera split-intein moiety.
  • Protein trans-splicing (PTS) occurs in vivo when split inteins interact due to their high affinity.
  • A new peptide bond is formed between N-term and C-term regions, reconstituting the large Protein of Interest.

Targeting virus particles to specific tissues: Virus Surface Modification

SPLITTERA Split Inteins mediate the covalent display of targeting proteins on the surface of a virus. Targeted viral particles allow restricted and tissue specific gene delivery.

  • Targeting protein (eg Ab) in fusion with one intein moiety is produced recombinantly in the host of choice.
  • Viral Particle is produced recombinantly in fusion with the other intein moiety in the appropriate host.
  • Protein trans-splicing (PTS) occurs when split inteins interact due to its high affinity for each other.
  • Virus surface is covalently decorated with the desired protein.