Loop assembly: a simple and open system for recursive fabrication of DNA circuit

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dc.catalogadorjlo
dc.contributor.authorPollak, Bernardo
dc.contributor.authorCerda, Ariel
dc.contributor.authorDelmans, Mihails
dc.contributor.authorÁlamos, Simón
dc.contributor.authorMoyano, Tomás
dc.contributor.authorWest, Anthony
dc.contributor.authorGutiérrez, Rodrigo A.
dc.contributor.authorPatron, Nicola J.
dc.contributor.authorFederici, Fernán
dc.contributor.authorHaseloff, Jim
dc.date.accessioned2024-01-29T17:48:28Z
dc.date.available2024-01-29T17:48:28Z
dc.date.issued2019
dc.description.abstractHigh-efficiency methods for DNA assembly have enabled the routine assembly of syntheticDNAs of increased size and complexity. However, these techniques require customization,elaborate vector sets or serial manipulations for the different stages of assembly. We have developed Loop assembly based on a recursive approach to DNA fabrication. Thesystem makes use of two Type IIS restriction endonucleases and corresponding vector sets forefficient and parallel assembly of large DNA circuits. Standardized level 0 parts can be assem-bled into circuits containing 1, 4, 16 or more genes by looping between the two vector sets.The vectors also contain modular sites for hybrid assembly using sequence overlap methods. Loop assembly enables efficient and versatile DNA fabrication for plant transformation. Weshow the construction of plasmids up to 16 genes and 38 kb with high efficiency (> 80%).We have characterized Loop assembly on over 200 different DNA constructs and validatedthe fidelity of the method by high-throughput Illumina plasmid sequencing. Our method provides a simple generalized solution for DNA construction with standardizedparts. The cloning system is provided under an OpenMTA license for unrestricted sharing andopen access.
dc.fuente.origenORCID-ene24
dc.identifier.doi10.1111/nph.15625
dc.identifier.issn0028-646X
dc.identifier.urihttp://dx.doi.org/10.1111/nph.15625
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/81011
dc.information.autorucFacultad de Ciencias Biológicas; Cerda Rojas, Ariel Patricio; 0009-0007-3040-4849; 250244
dc.information.autorucFacultad de Ciencias Biológicas; Moyano Yugovic, Tomás Custodio; S/I; 149778
dc.information.autorucFacultad de Ciencias Biológicas; Gutiérrez Ilabaca, Rodrigo Antonio; 0000-0002-5961-5005; 86782
dc.information.autorucFacultad de Ciencias Biológicas; Federici, Fernán; 0000-0001-9200-5383; 123357
dc.issue.numero1
dc.language.isoen
dc.nota.accesoContenido parcial
dc.pagina.final640
dc.pagina.inicio628
dc.revistaNew Phytologist
dc.rightsacceso restringido
dc.subject.ddc610
dc.subject.deweyMedicina y saludes_ES
dc.titleLoop assembly: a simple and open system for recursive fabrication of DNA circuit
dc.typeartículo
dc.volumen222
sipa.codpersvinculados250244
sipa.codpersvinculados149778
sipa.codpersvinculados86782
sipa.codpersvinculados123357
sipa.trazabilidadORCID;2024-01-15
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