Naslov
Interplay of recombination, plasmid stability, and CRISPR-Cas immunity in Escherichia coli

Autori
Radovčić, Marin ; Bolt, Edward L., Ivančić-Baće, Ivana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Cell Symposia CRISPR: From biology to technology and novel therapeutics / Pham, John ; Saxe, Jon ; Joung, Keith ; May, Andy - Online : Cell Press, Elsevier, 2017, 3-3

Skup
Cell Symposia CRISPR: From biology to technology and novel therapeutics

Mjesto i datum
Sitges, Španjolska, 22.10.2017. - 24.10.2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Cas1-Cas2, RecBCD, RecA, E. coli

Sažetak
CRISPR-Cas system is a prokaryotic adaptive immune system against foreign genetic elements. Immunity is acquired through insertion of small fragments of invader DNA into a CRISPR array. This process of fragment (spacer) insertion is called adaptation, and can be mediated solely by the protein complex Cas1-Cas2 (“naïve CRISPR adaptation”). In E. coli, naïve adaptation is helped by RecBCD enzyme to generate single-stranded DNA intermediates. We report here that cells lacking recD gene and transformed with Cas1-Cas2 expressing plasmid could not acquire new spacers, as expected, but that this corresponded with moderate loss of pCas1-Cas2. The adaptation could be re-established by deletion of recA, but the plasmid loss remained and the cell’s viability became reduced. Since RecA loading is constitutive in recD mutant, the presence of bound RecA onto 3’ ssDNA likely prevents reannealing of ssDNA fragments while at the same time it promotes recombination that generates unstable plasmid multimers.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA