TY - JOUR
T1 - PlasmidMaker is a versatile, automated, and high throughput end-to-end platform for plasmid construction
AU - Enghiad, Behnam
AU - Xue, Pu
AU - Singh, Nilmani
AU - Boob, Aashutosh Girish
AU - Shi, Chengyou
AU - Petrov, Vassily Andrew
AU - Liu, Roy
AU - Peri, Siddhartha Suryanarayana
AU - Lane, Stephan Thomas
AU - Gaither, Emily Danielle
AU - Zhao, Huimin
N1 - This work was supported by U.S. Department of Energy award DE-SC0018420. Any opinions, findings, and conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the U.S. Department of Energy. We thank Jonathan Chow Ning for providing the zeaxanthin plasmid template and helpful discussions about initial design of experiments. The online tool BioRender (biorender.com) was used to create Fig. ’s upper panel, Fig. , Fig. , and Supplementary Fig. , .
PY - 2022/12
Y1 - 2022/12
N2 - Plasmids are used extensively in basic and applied biology. However, design and construction of plasmids, specifically the ones carrying complex genetic information, remains one of the most time-consuming, labor-intensive, and rate-limiting steps in performing sophisticated biological experiments. Here, we report the development of a versatile, robust, automated end-to-end platform named PlasmidMaker that allows error-free construction of plasmids with virtually any sequences in a high throughput manner. This platform consists of a most versatile DNA assembly method using Pyrococcus furiosus Argonaute (PfAgo)-based artificial restriction enzymes, a user-friendly frontend for plasmid design, and a backend that streamlines the workflow and integration with a robotic system. As a proof of concept, we used this platform to generate 101 plasmids from six different species ranging from 5 to 18 kb in size from up to 11 DNA fragments. PlasmidMaker should greatly expand the potential of synthetic biology.
AB - Plasmids are used extensively in basic and applied biology. However, design and construction of plasmids, specifically the ones carrying complex genetic information, remains one of the most time-consuming, labor-intensive, and rate-limiting steps in performing sophisticated biological experiments. Here, we report the development of a versatile, robust, automated end-to-end platform named PlasmidMaker that allows error-free construction of plasmids with virtually any sequences in a high throughput manner. This platform consists of a most versatile DNA assembly method using Pyrococcus furiosus Argonaute (PfAgo)-based artificial restriction enzymes, a user-friendly frontend for plasmid design, and a backend that streamlines the workflow and integration with a robotic system. As a proof of concept, we used this platform to generate 101 plasmids from six different species ranging from 5 to 18 kb in size from up to 11 DNA fragments. PlasmidMaker should greatly expand the potential of synthetic biology.
UR - http://www.scopus.com/inward/record.url?scp=85130189437&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85130189437&partnerID=8YFLogxK
U2 - 10.1038/s41467-022-30355-y
DO - 10.1038/s41467-022-30355-y
M3 - Article
C2 - 35577775
AN - SCOPUS:85130189437
SN - 2041-1723
VL - 13
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 2697
ER -