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New method of DNA editing allows synthetic biologists to unlock secrets of a bacterial genome
Date:12/5/2013

tural products. This was the challenge that Zhao and colleagues hoped to address when they began their project. "Sequence analysis of bacterial genomes indicates that there are many cryptic or silent pathways that have not been discovered," Zhao said. " . . .they need the right signal to turn on expression of the whole gene cluster."

Several strategies have been employed to trick cells into activating their little-used, "cryptic" gene clusters, such as culturing bacteria in a variety of harsh conditions or inserting sets of genes from one species of bacteria into the genome of another species. These techniques involve labor-intensive trial and error, with no guarantee of success.

Zhao's group, rather than attempting to manipulate the environment, focused on reprogramming the control of gene expression within the cell. They used a genetic engineering method previously developed by Zhao's laboratory, called DNA assembler, to insert small sections of DNA between each gene in a cryptic gene cluster. The sections of DNA added were promoters, specialized regions that help control when and how much nearby genes are expressed. By adding the right promoters, Zhao and colleagues forced the cell to increase expression of every gene in the cluster.

What makes Zhao's strategy possible is the ability of the DNA assembler method to join many different fragments of DNA in a single step. Previous methods for DNA editing limited researchers to making a series of sequential changes; the number of experimental steps required to add a promoter to each gene in even a small cluster would have been prohibitive. In contrast, Zhao said, "we can actually build the whole cluster, so that gives us ultimate flexibility, because we can add different promoters," ensuring that every gene within the cluster is consistently activated.

For the study published in Nature Communications, Zhao and his coauthors modified a cryptic cluster of six genes fro
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Contact: Nicholas Vasi
nvasi@illinois.edu
Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Source:Eurekalert

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