Related Experiment Video
Updated: May 9, 2026
11:22
Automated Robotic Liquid Handling Assembly of Modular DNA Devices
Published on: December 1, 2017
Randomized BioBrick assembly: a novel DNA assembly method for randomizing and optimizing genetic circuits and
Sean C Sleight1, Herbert M Sauro
1Department of Bioengineering, University of Washington , Seattle, Washington 98195, United States.
ACS Synthetic Biology
|July 12, 2013
Summary
This study introduces a novel DNA assembly method for rapidly creating diverse genetic circuits and metabolic pathways. This approach enables efficient screening of numerous combinations to optimize biological functions and product yields.
Area of Science:
- Synthetic Biology
- Molecular Biology
- Metabolic Engineering
Background:
- Optimizing genetic circuits and metabolic pathways typically requires iterative construction or directed evolution.
- Screening large libraries of randomized genetic elements offers an alternative optimization strategy.
- A method to simultaneously randomize multiple components within a single assembly reaction is needed.
Purpose of the Study:
- To develop and demonstrate a new DNA assembly method for randomizing genetic circuits and metabolic pathways.
- To validate the method's efficacy in generating diverse and functional genetic constructs.
- To assess the impact of randomization on gene expression and metabolic product yield.
Main Methods:
- Developed a Gibson Assembly-based method to randomize modular DNA fragments (BioBricks) for genetic circuits.
- Assembled Cyan-Magenta-Yellow (CMY) three-gene circuits with independently randomized promoters, ribosome binding sites, and terminators.
- Adapted the method to randomize enzyme-coding sequences in the lycopene biosynthesis pathway.
Main Results:
- Successfully assembled CMY circuits with randomized parts; 20 out of 24 unique circuits were confirmed by sequencing.
- Observed a 200-fold range of gene expression levels above background in the randomized CMY circuits.
- Achieved a ~30% improvement in lycopene production compared to the highest-expressing polycistronic pathway.
Conclusions:
- The developed assembly method efficiently generates a vast number of unique genetic circuit and pathway combinations (nearly 20,000 possible with three randomized parts).
- This randomization approach significantly enhances screening capabilities for optimizing biological functions.
- The methodology holds potential for broad application in maximizing desired products from engineered circuits and pathways.
Related Concept Videos
03:36Genome Annotation and Assembly
The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
02:55Synthetic Biology
Synthetic biology is an interdisciplinary science that involves using principles from disciplines such as engineering, molecular biology, cell biology, and systems biology. It involves remodeling existing organisms from nature or constructing completely new synthetic organisms for applications such as protein or enzyme production, bioremediation, value-added macromolecule production, and the addition of desirable traits to crops, to name a few.
Golden rice
Golden rice is a genetically modified...
Golden rice
Golden rice is a genetically modified...
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
03:00Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.