Recombinase-mediated cassette exchange (RMCE) system for functional genomics studies in Mycoplasma mycoides
Open accessAbstract
BACKGROUND: We have previously established technologies enabling us to engineer the Mycoplasma mycoides genome while cloned in the yeast Saccharomyces cerevisiae, followed by genome transplantation into Mycoplasma capricolum recipient cells to produce M. mycoides with an altered genome. To expand the toolbox for genomic modifications, we designed a strategy based on the Cre/loxP-based Recombinase-Mediated Cassette Exchange (RMCE) system for functional genomics analyses. RESULTS: In this paper, we demonstrated replacement of an approximately 100 kb DNA segment of the M. mycoides genome with a synthetic DNA counterpart in two orientations. The function of the altered genomes was then validated by genome transplantation and phenotypic characterization of the transplanted cells. CONCLUSION: This method offers an easy and efficient way to manipulate the M. mycoides genome and will be a valuable tool for functional genomic studies, such as genome organization and minimization.
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JCVI-syn3.0 subclone 13-2 contains two heterospecific loxP sites that comprise a landing pad between genes 601 and 606.
“JCVI-syn3.0 subclone 13-2, referred to simply as JCVI-syn3.0 in this study, contains two heterospecific loxP sites that comprise a landing pad between genes 601 and 606 , which are adjacent in the fully-minimized JCVI-syn3.0 genome (Hutchison et al., 2016; Noskov et al., 2015).”
From Genetic requirements for cell division in a genomically minimal cell (2020) · cites this paper as Noskov et al., 2015
The Results section (‘Design of the Recombinase-Mediated Cassette Exchange’) describes a landing pad containing two heterospecific loxP sites. The paper describes its use in an M. mycoides genome, but our copy does not establish that JCVI-syn3.0 subclone 13-2 contains the pad or place it between genes 601 and 606. (LLM, conf=0.88)
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Direct Genome Transfer from Acholeplasma laidlawii to Yeast
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- Authors
- Noskov, V. N., Ma, L., Chen, S., & Chuang, R.-Y.
- Affiliations
- J. Craig Venter Institute (US)
- Corresponding author
- Chuang, Ray-Yuan
- ORCID
- Venue
- Biological Procedures Online
- Published
- 2015-03-02
- Volume
- 17
- Issue
- 1
- Pages
- 6-6
- ISSN
- 1480-9222 (ISSN-L)
- PMC
- PMC4359775
- OpenAlex
- W2123271834
- Cited by
- 12
- References
- 17
- Citations by year
- 2026: 1 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 2 · 2020: 2 · 2019: 1 · 2016: 3
- FWCI
- 0.43
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Biological Procedures Online (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Defense Advanced Research Projects Agency (HR0011)
- Advanced Research Projects Agency
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Immunology and Microbiology › Microbiology › Microbial infections and disease research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- recombinase-mediated cassette exchange, Mycoplasma mycoides, Cre-loxP system, genome editing, genome organization, functional genomics, Mycoplasma capricolum
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