A Modular Assembly Platform for Rapid Generation of DNA Constructs

作者：Akama Garren Elliot H; Joshi Nikhil S; Tammela Tuomas; Chang Gregory P; Wagner Bethany L; Lee Da Yae; Rideout William M III; Papagiannakopoulos Thales; Xue Wen; Jacks Tyler^*

来源：Scientific Reports, 2016, 6(1): 16836.

DOI：10.1038/srep16836

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.

• 出版日期2016-2-18