Part:BBa_K5142048

Click Virus

This part is a genetically engineered vaccinia virus (VACV) with modified A27L protein containing three 4-azido-L-phenylalanine residues close to its N-terminus. The azido groups serve as the sites for the click reaction of azido and dibenzocyclooctyne (DBCO) groups, which is the reason why this part is named “Click Virus”. Since A27L distributes on the surface of VACV and azido-DBCO click reaction can occur under physiological conditions without any catalyst, it is quite easy to link any customized DBCO-conjugated element onto the surface of Click Virus covalently via click reaction. Depending on the molecules linked, the modified Click Virus can be applied to a wide variety of biological and medical fields, such as targeting delivery of genes, oncolytic virus and vaccine carrier. In summary, Click Virus is a pluripotent chassis allowing rapid development of customized viral particles without further genetical engineering.

Usage and Biology

This part is derived from Orthopoxvirus vaccinia Tian Tan (AF095689.1) by mutating 3 codons in A27L gene to amber suppressor codons (TAG) and translating them to 4-azido-L-phenylalanine using an artificial translation system. We chose to mutate A27L to reduce the biosafety risk of Click Virus in use. Since the amber suppressor mutations are close to the N-terminus of A27L, premature termination of A27L translation will occur if the artificial translation system is not provided. Therefore, all the progeny of Click Virus after the first round of infection will be A27L-deficiency virus which will be trapped in the target cells due to impaired ability of dissemination./span>

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]