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

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

© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

Bibliographische Detailangaben
Veröffentlicht in:The New phytologist. - 1979. - 222(2019), 1 vom: 01. Apr., Seite 628-640
1. Verfasser: Pollak, Bernardo (VerfasserIn)
Weitere Verfasser: Cerda, Ariel, Delmans, Mihails, Álamos, Simón, Moyano, Tomás, West, Anthony, Gutiérrez, Rodrigo A, Patron, Nicola J, Federici, Fernán, Haseloff, Jim
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2019
Zugriff auf das übergeordnete Werk:The New phytologist
Schlagworte:Journal Article Research Support, Non-U.S. Gov't LoopDesigner Loop assembly OpenMTA Type IIS common syntax recursive assembly standardized DNA assembly unique nucleotide sequences (UNSs) DNA, Plant
Beschreibung
Zusammenfassung:© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.
High-efficiency methods for DNA assembly have enabled the routine assembly of synthetic DNAs 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. The system makes use of two Type IIS restriction endonucleases and corresponding vector sets for efficient and parallel assembly of large DNA circuits. Standardized level 0 parts can be assembled 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. We show 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 validated the fidelity of the method by high-throughput Illumina plasmid sequencing. Our method provides a simple generalized solution for DNA construction with standardized parts. The cloning system is provided under an OpenMTA license for unrestricted sharing and open access
Beschreibung:Date Completed 27.02.2020
Date Revised 09.01.2021
published: Print-Electronic
Citation Status MEDLINE
ISSN:1469-8137
DOI:10.1111/nph.15625