{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Shen K"],"funding":["NIGMS NIH HHS"],"pubmed_abstract":["Long single-stranded DNA (ssDNA) is a versatile molecular reagent with applications including RNA-guided genome engineering and DNA nanotechnology, yet its production is typically resource-intensive. We introduce a novel method utilizing an engineered <i>E. coli</i> \"helper\" strain and phagemid system that simplifies long ssDNA generation to a straightforward transformation and purification procedure. Our method obviates the need for helper plasmids and their associated contamination by integrating M13mp18 genes directly into the <i>E. coli</i> chromosome. We achieved ssDNA lengths ranging from 504 to 20,724 nucleotides with titers up to 250 μg/L following alkaline-lysis purification. The efficacy of our system was confirmed through its application in primary T cell genome modifications and DNA origami folding. The reliability, scalability, and ease of our approach promises to unlock new experimental applications requiring large quantities of long ssDNA."],"journal":["bioRxiv : the preprint server for biology"],"pagination":["2024.02.27.582394"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC10925301"],"repository":["biostudies-literature"],"pubmed_title":["Engineering an Escherichia coli strain for production of long single-stranded DNA."],"pmcid":["PMC10925301"],"funding_grant_id":["R35 GM125027","F32 GM147967"],"pubmed_authors":["Shen K","Flood JJ","Zhang Z","Ha A","Douglas SM","Dueber JE","Shy BR"],"additional_accession":[]},"is_claimable":false,"name":"Engineering an Escherichia coli strain for production of long single-stranded DNA.","description":"Long single-stranded DNA (ssDNA) is a versatile molecular reagent with applications including RNA-guided genome engineering and DNA nanotechnology, yet its production is typically resource-intensive. We introduce a novel method utilizing an engineered <i>E. coli</i> \"helper\" strain and phagemid system that simplifies long ssDNA generation to a straightforward transformation and purification procedure. Our method obviates the need for helper plasmids and their associated contamination by integrating M13mp18 genes directly into the <i>E. coli</i> chromosome. We achieved ssDNA lengths ranging from 504 to 20,724 nucleotides with titers up to 250 μg/L following alkaline-lysis purification. The efficacy of our system was confirmed through its application in primary T cell genome modifications and DNA origami folding. The reliability, scalability, and ease of our approach promises to unlock new experimental applications requiring large quantities of long ssDNA.","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Mar","modification":"2025-04-19T20:26:31.77Z","creation":"2025-04-19T20:26:31.77Z"},"accession":"S-EPMC10925301","cross_references":{"pubmed":["38464036"],"doi":["10.1101/2024.02.27.582394"]}}