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A Modular Assembly Platform for Rapid Generation of DNA Constructs

Traditional cloning methods have limitations on the number of DNA fragments that can be simultaneously manipulated, which dramatically slows the pace of molecular assembly. Here we describe GMAP, a Gibson assembly-based modular assembly platform consisting of a collection of promoters and genes, whi...

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Autores principales: Akama-Garren, Elliot H., Joshi, Nikhil S., Tammela, Tuomas, Chang, Gregory P., Wagner, Bethany L., Lee, Da-Yae, Rideout III, William M., Papagiannakopoulos, Thales, Xue, Wen, Jacks, Tyler
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4757859/
https://www.ncbi.nlm.nih.gov/pubmed/26887506
http://dx.doi.org/10.1038/srep16836
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author Akama-Garren, Elliot H.
Joshi, Nikhil S.
Tammela, Tuomas
Chang, Gregory P.
Wagner, Bethany L.
Lee, Da-Yae
Rideout III, William M.
Papagiannakopoulos, Thales
Xue, Wen
Jacks, Tyler
author_facet Akama-Garren, Elliot H.
Joshi, Nikhil S.
Tammela, Tuomas
Chang, Gregory P.
Wagner, Bethany L.
Lee, Da-Yae
Rideout III, William M.
Papagiannakopoulos, Thales
Xue, Wen
Jacks, Tyler
author_sort Akama-Garren, Elliot H.
collection PubMed
description Traditional cloning methods have limitations on the number of DNA fragments that can be simultaneously manipulated, which dramatically slows the pace of molecular assembly. Here we describe GMAP, a Gibson assembly-based modular assembly platform consisting of a collection of promoters and genes, which allows for one-step production of DNA constructs. GMAP facilitates rapid assembly of expression and viral constructs using modular genetic components, as well as increasingly complicated genetic tools using contextually relevant genomic elements. Our data demonstrate the applicability of GMAP toward the validation of synthetic promoters, identification of potent RNAi constructs, establishment of inducible lentiviral systems, tumor initiation in genetically engineered mouse models, and gene-targeting for the generation of knock-in mice. GMAP represents a recombinant DNA technology designed for widespread circulation and easy adaptation for other uses, such as synthetic biology, genetic screens, and CRISPR-Cas9.
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spelling pubmed-47578592016-02-26 A Modular Assembly Platform for Rapid Generation of DNA Constructs Akama-Garren, Elliot H. Joshi, Nikhil S. Tammela, Tuomas Chang, Gregory P. Wagner, Bethany L. Lee, Da-Yae Rideout III, William M. Papagiannakopoulos, Thales Xue, Wen Jacks, Tyler Sci Rep Article Traditional cloning methods have limitations on the number of DNA fragments that can be simultaneously manipulated, which dramatically slows the pace of molecular assembly. Here we describe GMAP, a Gibson assembly-based modular assembly platform consisting of a collection of promoters and genes, which allows for one-step production of DNA constructs. GMAP facilitates rapid assembly of expression and viral constructs using modular genetic components, as well as increasingly complicated genetic tools using contextually relevant genomic elements. Our data demonstrate the applicability of GMAP toward the validation of synthetic promoters, identification of potent RNAi constructs, establishment of inducible lentiviral systems, tumor initiation in genetically engineered mouse models, and gene-targeting for the generation of knock-in mice. GMAP represents a recombinant DNA technology designed for widespread circulation and easy adaptation for other uses, such as synthetic biology, genetic screens, and CRISPR-Cas9. Nature Publishing Group 2016-02-18 /pmc/articles/PMC4757859/ /pubmed/26887506 http://dx.doi.org/10.1038/srep16836 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Akama-Garren, Elliot H.
Joshi, Nikhil S.
Tammela, Tuomas
Chang, Gregory P.
Wagner, Bethany L.
Lee, Da-Yae
Rideout III, William M.
Papagiannakopoulos, Thales
Xue, Wen
Jacks, Tyler
A Modular Assembly Platform for Rapid Generation of DNA Constructs
title A Modular Assembly Platform for Rapid Generation of DNA Constructs
title_full A Modular Assembly Platform for Rapid Generation of DNA Constructs
title_fullStr A Modular Assembly Platform for Rapid Generation of DNA Constructs
title_full_unstemmed A Modular Assembly Platform for Rapid Generation of DNA Constructs
title_short A Modular Assembly Platform for Rapid Generation of DNA Constructs
title_sort modular assembly platform for rapid generation of dna constructs
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4757859/
https://www.ncbi.nlm.nih.gov/pubmed/26887506
http://dx.doi.org/10.1038/srep16836
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