A method for targeting a specified segment of DNA to a bacterial microorganelleOtoničar, Jan (Avtor) Hostnik, Maja (Avtor) Grundner, Maja (Avtor) Kostanjšek, Rok (Avtor) Gredar, Tajda (Avtor) Zorc, Maja (Avtor) Arsov, Zoran (Avtor) Podlesek, Zdravko (Avtor) Gostinčar, Cene (Avtor) Jakše, Jernej (Avtor) Busby, Steve J. W. (Avtor) Butala, Matej (Avtor) DNAphysical characterisationbiochemical characterisationEncapsulation of a selected DNA molecule in a cell has important implications for bionanotechnology. Non-viral proteins that can be used as nucleic acid containers include proteinaceous subcellular bacterial microcompartments (MCPs) that self-assemble into a selectively permeable protein shell containing an enzymatic core. Here, we adapted a propanediol utilization (Pdu) MCP into a synthetic protein cage to package a specified DNA segment in vivo, thereby enabling subsequent affinity purification. To this end, we engineered the LacI transcription repressor to be routed, together with target DNA, into the lumen of a Strep-tagged Pdu shell. Sequencing of extracted DNA from the affinity-isolated MCPs shows that our strategy results in packaging of a DNA segment carrying multiple LacI binding sites, but not the flanking regions. Furthermore, we used LacI to drive the encapsulation of a DNA segment containing operators for LacI and for a second transcription factor.20222023-01-10 14:28:11Članek v reviji143717UDK: 577ISSN pri članku: 0305-1048DOI: 10.1093/nar/gkac714COBISS_ID: 120251907sl