ArticleMicrobial cell factories2019
Efficient Cas9-based genome editing of Rhodobacter sphaeroides for metabolic engineering.
Article in Microbial cell factories, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
15 citing papers in PubMed, 38 citations in OpenAlex.
- ETfinder: harnessing conserved C-terminal tails of single-stranded DNA-binding proteins for mining and engineering RecET systems in non-model microbial chassis.Nucleic acids research · 2026Article
- Extracellular Metabolite Profiling in COMetabolites · 2026Article
- Genetic tools for engineering Zymomonas mobilis, Cereibacter sphaeroides and Novosphingobium aromaticivorans to improve production of bioenergy compounds.Microbial cell factories · 2025Review
- Systematic engineering to enhance valencene production in Rhodobacter sphaeroides.Bioresources and bioprocessing · 2025Article
- Optimization of Electroporation Conditions for Introducing Heterologous DNA intoJournal of microbiology and biotechnology · 2024Article
- Differential stability of bacterial photosynthetic apparatus ofFrontiers in microbiology · 2024Article
- Synthetic Biology Toolbox, Including a Single-Plasmid CRISPR-Cas9 System to Biologically Engineer the Electrogenic, Metal-Resistant BacteriumACS synthetic biology · 2022Article
- Characteristics and Application ofFrontiers in bioengineering and biotechnology · 2022Review
- Screening and engineering of high-activity promoter elements through transcriptomics and red fluorescent protein visualization inSynthetic and systems biotechnology · 2021Article
- CRISPR/Cas12a-mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus.Microbial biotechnology · 2021Article
- The transition of Rhodobacter sphaeroides into a microbial cell factory.Biotechnology and bioengineering · 2021Review
- An overview of anoxygenic phototrophic bacteria and their applications in environmental biotechnology for sustainable Resource recovery.Biotechnology reports (Amsterdam, Netherlands) · 2020Review
- Functional replacement of isoprenoid pathways in Rhodobacter sphaeroides.Microbial biotechnology · 2020Article
- CRISPR/Cas9-deaminase enables robust base editing in Rhodobacter sphaeroides 2.4.1.Microbial cell factories · 2020Article
- Growth-uncoupled isoprenoid synthesis inBiotechnology for biofuels · 2020Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
9 authors at 1 institution in 2 countries.
Funding
Abstract
backgroundRhodobacter sphaeroides is a metabolically versatile bacterium that serves as a model for analysis of photosynthesis, hydrogen production and terpene biosynthesis. The elimination of by-products formation, such as poly-β-hydroxybutyrate (PHB), has been an important metabolic engineering target for R. sphaeroides. However, the lack of efficient markerless genome editing tools for R. sphaeroides is a bottleneck for fundamental studies and biotechnological exploitation. The Cas9 RNA-guided DNA-endonuclease from the type II CRISPR-Cas system of Streptococcus pyogenes (SpCas9) has been extensively employed for the development of genome engineering tools for prokaryotes and eukaryotes, but not for R. sphaeroides.
resultsHere we describe the development of a highly efficient SpCas9-based genomic DNA targeting system for R. sphaeroides, which we combine with plasmid-borne homologous recombination (HR) templates developing a Cas9-based markerless and time-effective genome editing tool. We further employ the tool for knocking-out the uracil phosphoribosyltransferase (upp) gene from the genome of R. sphaeroides, as well as knocking it back in while altering its start codon. These proof-of-principle processes resulted in editing efficiencies of up to 100% for the knock-out yet less than 15% for the knock-in. We subsequently employed the developed genome editing tool for the consecutive deletion of the two predicted acetoacetyl-CoA reductase genes phaB and phbB in the genome of R. sphaeroides. The culturing of the constructed knock-out strains under PHB producing conditions showed that PHB biosynthesis is supported only by PhaB, while the growth of the R. sphaeroides ΔphbB strains under the same conditions is only slightly affected.
conclusionsIn this study, we combine the SpCas9 targeting activity with the native homologous recombination (HR) mechanism of R. sphaeroides for the development of a genome editing tool. We further employ the developed tool for the elucidation of the PHB production pathway of R. sphaeroides. We anticipate that the presented work will accelerate molecular research with R. sphaeroides.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.