An educational module to explore CRISPR technologies with a cell-free transcription-translation system
نویسندگان
چکیده
منابع مشابه
Rapid and Scalable Characterization of CRISPR Technologies Using an E. coli Cell-Free Transcription-Translation System.
CRISPR-Cas systems offer versatile technologies for genome engineering, yet their implementation has been outpaced by ongoing discoveries of new Cas nucleases and anti-CRISPR proteins. Here, we present the use of E. coli cell-free transcription-translation (TXTL) systems to vastly improve the speed and scalability of CRISPR characterization and validation. TXTL can express active CRISPR machine...
متن کاملImplementation and System Identification of a Phosphorylation-Based Insulator in a Cell-Free Transcription-Translation System
An outstanding challenge in the design of synthetic biocircuits is the development of a robust and efficient strategy for interconnecting functional modules. Recent work demonstrated that a phosphorylation-based insulator (PBI) implementing a dual strategy of high gain and strong negative feedback can be used as a device to attenuate retroactivity. This paper describes the implementation of suc...
متن کاملA versatile coupled cell-free transcription-translation system based on tobacco BY-2 cell lysates.
Cell-free protein synthesis is a powerful method for the high-throughput production of recombinant proteins, especially proteins that are difficult to express in living cells. Here we describe a coupled cell-free transcription-translation system based on tobacco BY-2 cell lysates (BYLs). Using a combination of fractional factorial designs and response surface models, we developed a cap-independ...
متن کاملPrototyping a valinomycin biosynthesis pathway within a cell-free transcription-translation (TX-TL) system
Many natural metabolites have antibacterial, antiviral, or anticancer effects and can be developed into new drugs. However, working with the microorganisms that produce these products can be challenging since they are not as well characterized as a model organism like Escherichia coli. In this paper, we investigate the potential for a cell-free transcription-translation (TX-TL) system to provid...
متن کاملTitle Prototyping 1,4-butanediol (BDO) biosynthesis pathway in a cell-free transcription-translation (TX-TL) system
Current methods for assembling metabolic pathways require a process of repeated trial and error and have a long design-build-test cycle. Further, it remains a challenge to precisely tune enzyme expression levels for maximizing target metabolite production. Recently it was shown that a cellfree transcriptional-translation system (TX-TL) can be used to rapidly prototype novel complex biocircuits ...
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ژورنال
عنوان ژورنال: Synthetic Biology
سال: 2019
ISSN: 2397-7000
DOI: 10.1093/synbio/ysz005