Single day construction of multi-gene circuits with 3G assembly

Title	Single day construction of multi-gene circuits with 3G assembly
Authors	Andrew D Halleran, Anandh Swaminathan and Richard M Murray
Source	2018 Synthetic Biology: Engineering, Evolution and Design (SEED) Conference
Abstract	The ability to rapidly design, build, and test prototypes is of key importance to every engineering discipline. DNA assembly often serves as a rate limiting step of the prototyping cycle for synthetic biology. Recently developed DNA assembly methods such as isothermal assembly and type IIS restriction enzyme systems take different approaches to accelerate DNA construction. We introduce a hybrid method, Golden Gate-Gibson (3G), that takes advantage of modular part libraries introduced by type IIS restriction enzyme systems and isothermal assembly's ability to build large DNA constructs in single pot reactions. Our method is highly efficient and rapid, facilitating construction of entire multi-gene circuits in a single day. Additionally, 3G allows generation of variant libraries enabling efficient screening of different possible circuit constructions. We characterize the efficiency and accuracy of 3G assembly for various construct sizes, and demonstrate 3G by characterizing variants of an inducible cell-lysis circuit.
Type	Conference paper
URL	https://www.biorxiv.org/content/early/2018/02/10/260851
DOI
Tag	hsm18-seed
ID	2018a
Funding	DARPA BioCon
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