Details

Autor(en) / Beteiligte

• Qazi, Shefah
• Miettinen, Heini M.
• Wilkinson, Royce A.
• McCoy, Kimberly
• Douglas, Trevor
• Wiedenheft, Blake

Titel

Programmed Self-Assembly of an Active P22-Cas9 Nano Carrier System

Ist Teil von

• Molecular pharmaceutics, 2016-02, Vol.13 (3), p.1191-1196

Erscheinungsjahr

2016

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• C lustered R egularly I nterspaced S hort P alindromic R epeats (CRISPR) RNA-guided endonucleases are powerful new tools for targeted genome engineering. These nucleases provide an efficient and precise method for manipulating eukaryotic genomes; however, delivery of these reagents to specific cell-types remains challenging. Virus-like particles (VLPs) derived from bacteriophage P22, are robust supramolecular protein cage structures with demonstrated utility for cell type-specific delivery of encapsulated cargos. Here, we genetically fuse Cas9 to a truncated form of the P22 scaffold protein, which acts as a template for capsid assembly as well as a specific encapsulation signal for Cas9. Our results indicate that Cas9 and a single-guide RNA are packaged inside the P22 VLP and activity assays indicate that this RNA-guided endonuclease is functional for sequence-specific cleavage of dsDNA targets. This work demonstrates the potential for developing P22 as a delivery vehicle for cell specific targeting of Cas9.