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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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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. |
format | Online Article Text |
id | pubmed-4757859 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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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