A method for targeting a specified segment of DNA to a bacterial microorganelle

Otoničar, Jan (avtor), Hostnik, Maja (avtor), Grundner, Maja (avtor), Kostanjšek, Rok (avtor), Gredar, Tajda (avtor), Garvas, Maja (avtor), Arsov, Zoran (avtor), Podlesek, Zdravko (avtor), Gostinčar, Cene (avtor), Jakše, Jernej (avtor), Busby, Steve J. W. (avtor), Butala, Matej (avtor)

Encapsulation 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.

• Jezik: Angleški jezik
• Ključne besede: DNA, physical characterisation, biochemical characterisation
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2022
• Št. strani: 12 str.
• Številčenje: Vol. 50, no. 19, art. e113
• PID: 20.500.12556/RUL-143717
• UDK: 577
• ISSN pri članku: 0305-1048
• DOI: 10.1093/nar/gkac714
• COBISS.SI-ID: 120251907
• Datum objave v RUL: 10.01.2023

Gradivo je del revije

• Naslov: Nucleic acids research
• Skrajšan naslov: Nucleic acids res.
• Založnik: Oxford University Press, Nucleic Acids Research
• ISSN: 0305-1048
• COBISS.SI-ID: 3269647

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: bakterijski mikrorazdelek Pdu, Twin-Strep-tag, represor LacI, molekularna biologija, biokemija