Super Recombinator (SuRe): Anin vivorecombination system for scalable and efficient transgene assembly at a single genomic locus DOI Creative Commons
Junjie Luo, Cheng Huang, Caitlin Taylor

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: April 15, 2025

Abstract The capacity to engineer organisms with multiple transgenic components is crucial synthetic biology and basic research. For the former field, allow creation of novel biological functions; for latter, such provide potent means dissecting complex pathways. However, size limitations a single transgenesis event challenges associated assembly DNA fragments hinder efficient integration transgenes. To overcome these hurdles, here we introduce building block design termed an integrated genetic array (IGA), which incorporates all into locus prevent their separation during manipulations. Since natural recombination rate genes located in same near zero, construct IGAs developed Super Recombinator (SuRe) system, uses CRISPR/Cas9, alone or combination site-specific serine recombinases, vivo transgene at genomic locus. SuRe effectively doubles number elements assembled each round, exponentially accelerating IGA construction. By preventing elements, greatly reduces screening burdens, as validated through studies Drosophila melanogaster Caenorhabditis elegans . optimize SuRe, compared CRISPR/Cas9-induced homology-directed using various recombinases. Optimized versions achieved efficiency fidelity values theoretical maxima allowed generation recombinant products up 4.2 Mbp Using created fruit flies 12 fluorescence voltage imaging neural activity precisely defined cell-types. Mathematical modeling scalability large assemblies showed that times gene workloads respectively scale logarithmically linearly transgenes, both major improvements over conventional approaches. Overall, enables individual loci, chromosomal scale.

Language: Английский

Genetic circuits in synthetic biology: broadening the toolbox of regulatory devices DOI Creative Commons
Marik M. Müller, Katja M. Arndt, Stefan A. Hoffmann

et al.

Frontiers in Synthetic Biology, Journal Year: 2025, Volume and Issue: 3

Published: March 7, 2025

Devices sensing inputs and generating outputs are fundamental regulatory units, as such the basis of more complex networks. We provide an overview devices used building blocks in synthetic biology, how genetic circuitry is being constructed from them. first comprehensively explore operating at different levels gene regulation, with action modes on DNA sequence, to transcriptional, translational post-translational control. then discuss design principles constructing circuits basic addressing challenges orthogonality, context-dependence, noise, complexity. present examples circuitry, including bistable switches, logic gates, signal amplification, memory for biocomputation. How artificial can be useful real-life applications illustrated bioproduction, living therapeutics, biosafety. Our aim a comprehensive toolbox profound understanding their potential diverse applications.

Language: Английский

Citations

0
À l’origine du vivant, la différence sans hiérarchie(s) DOI Creative Commons
Jean‐Christophe Pagès

médecine/sciences, Journal Year: 2025, Volume and Issue: 41(3), P. 282 - 285

Published: March 1, 2025

Citations

0
Expediting genome synthesis of Corynebacterium glutamicum with an artificial chromosome vector DOI
Zhanhua Zhang,

Hong Pan,

Zebin Li

et al.

Trends in biotechnology, Journal Year: 2025, Volume and Issue: unknown

Published: March 1, 2025

Language: Английский

Citations

0
Super Recombinator (SuRe): Anin vivorecombination system for scalable and efficient transgene assembly at a single genomic locus DOI Creative Commons
Junjie Luo, Cheng Huang, Caitlin Taylor

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: April 15, 2025

Abstract The capacity to engineer organisms with multiple transgenic components is crucial synthetic biology and basic research. For the former field, allow creation of novel biological functions; for latter, such provide potent means dissecting complex pathways. However, size limitations a single transgenesis event challenges associated assembly DNA fragments hinder efficient integration transgenes. To overcome these hurdles, here we introduce building block design termed an integrated genetic array (IGA), which incorporates all into locus prevent their separation during manipulations. Since natural recombination rate genes located in same near zero, construct IGAs developed Super Recombinator (SuRe) system, uses CRISPR/Cas9, alone or combination site-specific serine recombinases, vivo transgene at genomic locus. SuRe effectively doubles number elements assembled each round, exponentially accelerating IGA construction. By preventing elements, greatly reduces screening burdens, as validated through studies Drosophila melanogaster Caenorhabditis elegans . optimize SuRe, compared CRISPR/Cas9-induced homology-directed using various recombinases. Optimized versions achieved efficiency fidelity values theoretical maxima allowed generation recombinant products up 4.2 Mbp Using created fruit flies 12 fluorescence voltage imaging neural activity precisely defined cell-types. Mathematical modeling scalability large assemblies showed that times gene workloads respectively scale logarithmically linearly transgenes, both major improvements over conventional approaches. Overall, enables individual loci, chromosomal scale.

Language: Английский

Citations

0