{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Hao W"],"funding":["Priority Academic Program Development of Jiangsu Higher Education Institutions","National Natural Science Foundation of China","Higher Education Discipline Innovation Project","Natural Science Foundation of Jiangsu Province"],"pagination":["14395-14409"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9749471"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["13(48)"],"pubmed_abstract":["The functionally evolved bacterial chassis is of great importance to manufacture a group of assorted high value-added chemicals, from small molecules to biologically active macromolecules. However, the current evolution frameworks are less efficienct in generating <i>in vivo</i> genomic diversification because of insufficient tunability, rendering limited evolution spacing for chassis. Here, an engineered genomic diversification platform (CRISPR-ABE8e-CDA-nCas9) leveraging a programmable dual-deaminases base editor was fabricated for rapidly evolving bacterial chassis. The dual-base editor was constructed by reprogramming the CRISPR array, nCas9, and cytidine and adenosine deaminase, enabling single or multiple base conversion at the genomic scale by simultaneous C-to-T and A-to-G conversion <i>in vivo</i>. Employing titration of the Cas-deaminase fusion protein, the platform enabled editing any pre-defined genomic loci with tunable conversion efficiency and editable window, generating a repertoire of mutants with highly diversified genomic sequences. Leveraging the genomic diversification platform, we successfully evolved the nisin-resistant capability of <i>Bacillus subtilis</i> through directed evolution of the subunit of lantibiotic ATP-binding cassette. Therefore, our work provides a portable and programmable genomic diversification platform, which is promising to expedite the fabrication of high-performance and robust bacterial chassis used in the development of biomanufacturing and biopharmaceuticals."],"journal":["Chemical science"],"pubmed_title":["Construction and application of an efficient dual-base editing platform for <i>Bacillus subtilis</i> evolution employing programmable base conversion."],"pmcid":["PMC9749471"],"funding_grant_id":["111-2-06","21878125","32171420","BK20181206"],"pubmed_authors":["Han L","Cui W","Hao W","Suo F","Cheng Z","Zhou Z"],"additional_accession":[]},"is_claimable":false,"name":"Construction and application of an efficient dual-base editing platform for <i>Bacillus subtilis</i> evolution employing programmable base conversion.","description":"The functionally evolved bacterial chassis is of great importance to manufacture a group of assorted high value-added chemicals, from small molecules to biologically active macromolecules. However, the current evolution frameworks are less efficienct in generating <i>in vivo</i> genomic diversification because of insufficient tunability, rendering limited evolution spacing for chassis. Here, an engineered genomic diversification platform (CRISPR-ABE8e-CDA-nCas9) leveraging a programmable dual-deaminases base editor was fabricated for rapidly evolving bacterial chassis. The dual-base editor was constructed by reprogramming the CRISPR array, nCas9, and cytidine and adenosine deaminase, enabling single or multiple base conversion at the genomic scale by simultaneous C-to-T and A-to-G conversion <i>in vivo</i>. Employing titration of the Cas-deaminase fusion protein, the platform enabled editing any pre-defined genomic loci with tunable conversion efficiency and editable window, generating a repertoire of mutants with highly diversified genomic sequences. Leveraging the genomic diversification platform, we successfully evolved the nisin-resistant capability of <i>Bacillus subtilis</i> through directed evolution of the subunit of lantibiotic ATP-binding cassette. Therefore, our work provides a portable and programmable genomic diversification platform, which is promising to expedite the fabrication of high-performance and robust bacterial chassis used in the development of biomanufacturing and biopharmaceuticals.","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Dec","modification":"2026-05-27T23:03:22.435Z","creation":"2024-11-10T03:45:09.651Z"},"accession":"S-EPMC9749471","cross_references":{"pubmed":["36545152"],"doi":["10.1039/d2sc05824c"]}}