ArticleNucleic acids research2020
Rapid and assured genetic engineering methods applied to Acinetobacter baylyi ADP1 genome streamlining.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 28 citations in OpenAlex.
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- Silica Nanoparticles Block Natural Genetic Transformation inACS omega · 2025Article
- Engineering microbial consortia for mixed plastic upcycling.Nature communications · 2025Article
- Genome evolution ofApplied and environmental microbiology · 2025Article
- A-to-I mRNA editing recodes hundreds of genes in dozens of species and produces endogenous protein isoforms in bacteria.Nucleic acids research · 2025Article
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- Evolution recovers the fitness of Acinetobacter baylyi strains with large deletions through mutations in deletion-specific targets and global post-transcriptional regulators.PLoS genetics · 2024Article
- Engineering cell morphology by CRISPR interference in Acinetobacter baylyi ADP1.Microbial biotechnology · 2022Article
- Article
- Bioelectricity generation using long-term operated biocathode: RFLP based microbial diversity analysis.Biotechnology reports (Amsterdam, Netherlands) · 2021Article
- EngineeringMetabolic engineering communications · 2021Article
- Defensive hypervariable regions confer superinfection exclusion in microviruses.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
One goal of synthetic biology is to improve the efficiency and predictability of living cells by removing extraneous genes from their genomes. We demonstrate improved methods for engineering the genome of the metabolically versatile and naturally transformable bacterium Acinetobacter baylyi ADP1 and apply them to a genome streamlining project. In Golden Transformation, linear DNA fragments constructed by Golden Gate Assembly are directly added to cells to create targeted deletions, edits, or additions to the chromosome. We tested the dispensability of 55 regions of the ADP1 chromosome using Golden Transformation. The 18 successful multiple-gene deletions ranged in size from 21 to 183 kb and collectively accounted for 23.4% of its genome. The success of each multiple-gene deletion attempt could only be partially predicted on the basis of an existing collection of viable ADP1 single-gene deletion strains and a new transposon insertion sequencing (Tn-Seq) dataset that we generated. We further show that ADP1's native CRISPR/Cas locus is active and can be retargeted using Golden Transformation. We reprogrammed it to create a CRISPR-Lock, which validates that a gene has been successfully removed from the chromosome and prevents it from being reacquired. These methods can be used together to implement combinatorial routes to further genome streamlining and for more rapid and assured metabolic engineering of this versatile chassis organism.
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