Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2021
Erindi/veggspjald / Talk/poster E69
Höfundar / Authors: Björn Þór Aðalsteinsson (1), Þórdís Kristjánsdóttir (1,2), Hörður Guðmundsson (1), William Merre (3), Alexandra Helleux (4), Julia Dusaucy (5), Mathilde Tourigny (4), Ólafur Friðjónsson (1), Guðmundur Óli Hreggviðsson (1,2)
Starfsvettvangur / Affiliations: 1. Matís, Reykjavík, Ísland; 2. Háskóli Íslands, Reykjavík, Ísland; 3. Aix-Marseille Université, Marseille, Frakkland; 4. Université de Strasbourg, Strassborg, Frakkland; 5. Université Pierre et Marie Curie, París, Frakkland
Kynnir / Presenter: Björn Aðalsteinsson
Genome editing systems for thermophilic organisms are relatively ill advanced – their use is often labor intensive and time consuming. We sought to develop an efficient Cas9 based genome editing system for thermophiles. We identified Cas9 and associated crRNA and tracrRNA in a thermophilic bacterium and designed a gRNA. The Cas9:gRNA pair displays targetable nuclease activity up to 65°C in vitro and a 5′-NNNNGNMA PAM specificity. We constructed a plasmid for their delivery and expression in thermophilic bacteria. Using the system we have generated knock-out mutants of the extreme thermophiles Thermus thermophilus and Rhodothermus marinus, and we are currently testing the system in a thermophilic anaerobe, Thermoanaerobacterium stain AK17. In T. thermophilus, cultured at 65°C, the system displayed 90% efficiency (Adalsteinsson et al 2021 Scientific Reports, 11:9586, doi.org/10.1038/s41598-021-89029-2). The system may facilitate genome editing efforts in thermophiles in general, and to that end it has been made available via the plasmid repository Addgene.