Development of a CRISPR/Cas12a-facilitated fluorescent aptasensor for sensitive detection of small molecules DOI
Ling Li, Menglei Li, Sai Wang

et al.

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 280, P. 136041 - 136041

Published: Sept. 26, 2024

Language: Английский

Discovery, characterization, and application of chromosomal integration sites for stable heterologous gene expression in Rhodotorula toruloides DOI
Haojie Xu, Lei Shi,

Aashutosh Girish Boob

et al.

Metabolic Engineering, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

Citations

0
CnRed: Efficient, Marker-free Genome Engineering of Cupriavidus necator H16 by Adapted Lambda Red Recombineering DOI Creative Commons

Simon Arhar,

Johanna Pirchner,

Holly Stolterfoht

et al.

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 24, 2025

Due to its ability utilize carbon dioxide, native intracellular accumulation of bioplastic precursors, and a high protein content, the bacterium Cupriavidus necator offers potential solutions for social problems tackled by modern biotechnology. Yet, engineering high-performing chemolithotrophic production strains has so far been hindered lack adequate genome editing methods. In this work we present establishment lambda Red recombineering system use in H16. combination with electroporation as DNA delivery system, it enables an efficient fast gene deletion methodology utilizing either suicide plasmids or, first time, linear PCR product. The novel was validated modification three different genomic loci and, proof-of-concept, ultimately utilized stable integration Escherichia coli phytase appA into phaC1 locus. A Cre/loxP further enabled marker recycling. minimal transformation protocol robust, freedom-to-operate synthetic biology tool increasingly important bacterial host. This approach simplifies accelerates C. is expected significantly enhance future strain development efforts.

Language: Английский

Citations

0
SHIP identifies genomic safe harbors in eukaryotic organisms using genomic general feature annotation DOI Creative Commons
Matheus de Castro Leitão, Lília I. L. Cabral, Luíza Cesca Piva

et al.

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: Feb. 28, 2025

Integrating foreign genes into loci, allowing their transcription without affecting endogenous gene expression, is the desirable strategy in genomic engineering. However, these known as safe harbors (GSHs), have been mainly identified by empirical methods. Furthermore, most prominent available GSHs are localized within regions of high density, raising concerns about unstable expression. As synthetic biology moving towards investigating polygenic modules rather than single genes, there an increasing demand for tools to identify systematically. To expand GSH repertoire, we present SHIP, algorithm designed detect potential eukaryotes. Using chassis organism Saccharomyces cerevisiae, five were experimentally curated based on data from DNA sequencing, stability, flow cytometry, qPCR, electron microscopy, RT-qPCR, and RNA-Seq assays. Our study places SHIP a valuable tool providing list promising candidates assist experimental assessment eukaryotic organisms with annotated genomes.

Language: Английский

Citations

0
Recent advances in CRISPR-Cas system for Saccharomyces cerevisiae engineering DOI

Xinxin Wu,

Xiaowen Wan, Hongbin Yu

et al.

Biotechnology Advances, Journal Year: 2025, Volume and Issue: 81, P. 108557 - 108557

Published: March 12, 2025

Language: Английский

Citations

0
Advancements in mammalian display technology for therapeutic antibody development and beyond: current landscape, challenges, and future prospects DOI Creative Commons
Peter Slavny,

Manjunath Hegde,

Achim Doerner

et al.

Frontiers in Immunology, Journal Year: 2024, Volume and Issue: 15

Published: Sept. 24, 2024

The evolving development landscape of biotherapeutics and their growing complexity from simple antibodies into bi- multi-specific molecules necessitates sophisticated discovery engineering platforms. This review focuses on mammalian display technology as a potential solution to the pressing challenges in biotherapeutic development. We provide comparative analysis with established methodologies, highlighting key aspects technology, including genetic engineering, construction libraries, its pivotal role hit selection and/or developability engineering. delves mechanisms underpinning developability-driven via broader implications. Applications beyond antibody are also explored, alongside advancements towards function-first screening technologies, precision genome AI/ML-enhanced situating them context display. Overall, provides comprehensive overview current landscape, underscores expansive for development, addresses critical full realisation this potential, examines advances related disciplines that might impact future application technologies.

Language: Английский

Citations

2
Development of a CRISPR/Cas12a-facilitated fluorescent aptasensor for sensitive detection of small molecules DOI
Ling Li, Menglei Li, Sai Wang

et al.

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 280, P. 136041 - 136041

Published: Sept. 26, 2024

Language: Английский

Citations

0