biostudies-literatureQazi SAmerican Heart AssociationAmgenMJ Murdock Charitable TrustOffice of Experimental Program to Stimulate Competitive ResearchNational Institute of General Medical SciencesNIGMS NIH HHSNational Science Foundation1191-6https://www.ebi.ac.uk/biostudies/studies/S-EPMC7734702biostudies-literatureUnknown13(3)Clustered Regularly Interspaced Short Palindromic Repeats (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 inMolecular pharmaceuticsProgrammed Self-Assembly of an Active P22-Cas9 Nanocarrier System.PMC7734702P20GM103500R01 GM108888P20 GM103500R01GM108888EPS-110134BMAT- 1507282McCoy KWilkinson RAQazi SWiedenheft BMiettinen HMDouglas TfalseProgrammed Self-Assembly of an Active P22-Cas9 Nanocarrier System.Clustered Regularly Interspaced Short Palindromic Repeats (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 in2016-01-01T00:00:00Z2016 Mar2025-04-22T02:08:02.388Z2021-02-20T10:54:11ZS-EPMC77347022689483610.1021/acs.molpharmaceut.5b00822