| Title: |
Integrase-On-Demand : bioprospecting integrases for targeted genomic insertion of genetic cargo |
| Authors: |
McClain, Hannah M; Lowrey, Lillian C; Quinto, Laura B; Torrance, Ellis L; Gagliano, Tomas R; Isaacs, Farren J; Schoeniger, Joseph S; Williams, Kelly P; Mageeney, Catherine M |
| Contributors: |
U.S. Department of Energy; Genomic Science Program, Office of Biological and Environmental Research; Secure Biosystems Design Science Focus Area, Intrinsic Control for Genome and Transcriptome Editing in Communities; InCoGenTEC; Sandia National Laboratories Laboratory Directed Research and Development; Office of Science; WDTS; Science Undergraduate Laboratory Internships Program; SULI; Department of Energy Office of Science BER |
| Source: |
Nucleic Acids Research ; volume 54, issue 4 ; ISSN 0305-1048 1362-4962 |
| Publisher Information: |
Oxford University Press (OUP) |
| Publication Year: |
2026 |
| Description: |
Integrases serve as powerful biotechnology tools that catalyze recombination at specific DNA sequences (att sites) and facilitate chromosomal integration of gene cargos transferred into cells. Given that genomes often lack the attB integration sites recognized by frequently utilized integrases, integrase technology has largely been restricted to genetic engineering of model organisms into which attB sites can be synthetically introduced. To enable single-step site-specific integrase-mediated genome editing in a broad spectrum of prokaryotes, we have devised the Integrase-On-Demand (IOD) method. IOD systematically identifies integrases, within bacteria and archaea, that can integrate into available attB sites in any target prokaryote. Computational results show that diverse bacteria generally have multiple potentially useable native attB sites for novel integrases. We confirmed the functionality of predicted integrase and attB pairs for mediating site-specific integration of heterologous DNA into the genomes of Pseudomonas putida S12 and KT2440 and Synechococcus elongatus UTEX 2973, measuring efficiency of integration using nonreplicating vectors. By eliminating the requirement to introduce non-native attB sites into the target genome, IOD may, when suitable transformation methods exist, allow facile genome integration of large constructs in nonmodel and possibly nonculturable bacteria. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1093/nar/gkag106 |
| Availability: |
https://doi.org/10.1093/nar/gkag106; https://academic.oup.com/nar/article-pdf/54/4/gkag106/66815207/gkag106.pdf |
| Rights: |
https://creativecommons.org/licenses/by/4.0/ |
| Accession Number: |
edsbas.226D572D |
| Database: |
BASE |