{"entity": "publication", "iuid": "2261d1c8b14b49248551a54eec077de5", "timestamp": "2026-08-22T06:57:14.265Z", "links": {"self": {"href": "https://publications-affiliated.scilifelab.se/publication/2261d1c8b14b49248551a54eec077de5.json"}, "display": {"href": "https://publications-affiliated.scilifelab.se/publication/2261d1c8b14b49248551a54eec077de5"}}, "title": "Inducible CRISPR/Cas9 Allows for Multiplexed and Rapidly Segregated Single-Target Genome Editing in Synechocystis Sp. PCC 6803.", "authors": [{"family": "Cengic", "given": "Ivana", "initials": "I"}, {"family": "Ca\u00f1adas", "given": "In\u00e9s C", "initials": "IC"}, {"family": "Minton", "given": "Nigel P", "initials": "NP", "orcid": "0000-0002-9277-1261", "researcher": {"href": "https://publications-affiliated.scilifelab.se/researcher/248e94d1f9574995a9ddf3beb3e84295.json"}}, {"family": "Hudson", "given": "Elton P", "initials": "EP", "orcid": "0000-0003-1899-7649", "researcher": {"href": "https://publications-affiliated.scilifelab.se/researcher/3b1c894d4c8b48569b9879d0342b91dd.json"}}], "type": "journal article", "published": "2022-09-16", "journal": {"title": "ACS Synth. Biol.", "issn": "2161-5063", "volume": "11", "issue": "9", "pages": "3100-3113", "issn-l": "2161-5063"}, "abstract": "Establishing various synthetic biology tools is crucial for the development of cyanobacteria for biotechnology use, especially tools that allow for precise and markerless genome editing in a time-efficient manner. Here, we describe a riboswitch-inducible CRISPR/Cas9 system, contained on a single replicative vector, for the model cyanobacterium Synechocystis sp. PCC 6803. A theophylline-responsive riboswitch allowed tight control of Cas9 expression, which enabled reliable transformation of the CRISPR/Cas9 vector intoSynechocystis. Induction of the CRISPR/Cas9 mediated various types of genomic edits, specifically deletions and insertions of varying size. The editing efficiency varied depending on the target and intended edit; smaller edits performed better, reaching, e.g., 100% for insertion of a FLAG-tag onto rbcL. Importantly, the single-vector CRISPR/Cas9 system mediated multiplexed editing of up to three targets in parallel inSynechocystis. All single-target and several double-target mutants were also fully segregated after the first round of induction. Lastly, a vector curing system based on the nickel-inducible expression of the toxic mazF (from Escherichia coli) was added to the CRISPR/Cas9 vector. This inducible system allowed for curing of the vector in 25-75% of screened colonies, enabling edited mutants to become markerless.", "doi": "10.1021/acssynbio.2c00375", "pmid": "35969224", "labels": [], "xrefs": [{"db": "pmc", "key": "PMC9486961"}], "notes": [], "created": "2026-08-20T08:13:54.138Z", "modified": "2026-08-20T08:45:52.187Z"}