ArticleScientific reports2016
One step engineering of the small-subunit ribosomal RNA using CRISPR/Cas9.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Protein-only centromeric chromatin assembly streamlines human artificial chromosome formation.bioRxiv : the preprint server for biology · 2026Article
- Rapid assembly of functional modules for generating human artificial chromosome constructs compatible with epigenetic centromere seeding.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2025Article
- Rapid assembly of functional modules for generating human artificial chromosome constructs compatible with epigenetic centromere seeding.bioRxiv : the preprint server for biology · 2025Article
- Synthetic collinearization of a chromosome segment enables genetic dissection of incompatibility between distant genera.bioRxiv : the preprint server for biology · 2025Article
- The Transformation Experiment of Frederick Griffith I: Its Narrowing and Potential for the Creation of Novel Microorganisms.Bioengineering (Basel, Switzerland) · 2025Review
- A toolbox for manipulating the genome of the major goat pathogen,Microbiology (Reading, England) · 2024Article
- Building synthetic chromosomes from natural DNA.Nature communications · 2023Article
- Evolution of a minimal cell.Nature · 2023Article
- Gene editing tools for mycoplasmas: references and future directions for efficient genome manipulation.Frontiers in microbiology · 2023Review
- Genetic basis of a spontaneous mutation's expressivity.Genetics · 2022Article
- Budding yeast as a factory to engineer partial and complete microbial genomes.Current opinion in systems biology · 2020Review
- Engineering the Translational Machinery for Biotechnology Applications.Molecular biotechnology · 2020Review
- Polar Effects of Transposon Insertion into a Minimal Bacterial Genome.Journal of bacteriology · 2019Article
Corrections and comments
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Authors and funding
12 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bacteria are indispensable for the study of fundamental molecular biology processes due to their relatively simple gene and genome architecture. The ability to engineer bacterial chromosomes is quintessential for understanding gene functions. Here we demonstrate the engineering of the small-ribosomal subunit (16S) RNA of Mycoplasma mycoides, by combining the CRISPR/Cas9 system and the yeast recombination machinery. We cloned the entire genome of M. mycoides in yeast and used constitutively expressed Cas9 together with in vitro transcribed guide-RNAs to introduce engineered 16S rRNA genes. By testing the function of the engineered 16S rRNA genes through genome transplantation, we observed surprising resilience of this gene to addition of genetic elements or helix substitutions with phylogenetically-distant bacteria. While this system could be further used to study the function of the 16S rRNA, one could envision the "simple" M. mycoides genome being used in this setting to study other genetic structures and functions to answer fundamental questions of life.
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