A proof-of-concept study involving the use of CRISPR and CRAGE technologies could make it easier to identify microbial secondary metabolites that are not essential for growth yet essential for survival.
The effectiveness of the tool for editing genomes has historically been limited due to the lack of robust tools. CRAGE is a technique that can be used to integrate large genes into diverse organisms.
Researchers can use the combination of CRAGE and CRISPR to study genes. The researchers used CRAGE-CRISPR to discover active secondary metabolites and help them understand the functions of the biosynthetic gene clusters.
There are secondary compounds that are produced in response to hardship. There's still so much we don't know about them, but they're the foundation for a lot of vital products.
BGCs that produce secondary metabolites can't be activated in a lab environment, so it can be difficult to gain access to their power. CRAGE was able to overcome this obstacle in the year 2019. The power is going to grow by combining it with a new tool.
Researchers can now use any microbe as a factory for producing chemical compounds of interest if they use CRAGE. JGI users can use CRAGE to domesticate target microbes.
There is a fatal photorhabdus luminescens. The host is quickly killed by the toxins that are released into the insect's bloodstream. Understanding how the work of P. luminescens could lead to new tools for controlling pests.
It's hard to identify which pathway corresponds to which secondary metabolite inbacteria. It's important to find a vehicle that will allow researchers to remove or modify genes and assess how these changes affect function.
CRAGE allows for the transfer of BGCs from one organisms to another through a landing pad that contains a cre recombinase gene and mutually exclusive lox sites. This process allows researchers to identify strains that can produce secondary metabolites in a lab environment.
A point of entry is also offered. Researchers at the JGI were able to monitor loss and gain-of-function with the help of the CRISPR technology. As genes are edited, the study shows peaks and valleys in secondary metabolites. The pair proved to confirm the enhanced production of 22 metabolites from six biosynthetic genes. There was a previously uncharacterized biosyntheticgene cluster that produced one of those. The work by JGI researchers was published in a journal.
Understanding how P. luminescens uses insects for fuel could lead to further agricultural applications for pest control.
It is possible that the impact of the pair will be far-reaching. The scientific community's understanding of how secondary metabolite are produced and how to leverage their powers in agriculture, pharmaceuticals, and beyond could be improved by the compatibility of CRAGE and CRISPR.
More information: Jing Ke et al, CRAGE-CRISPR facilitates rapid activation of secondary metabolite biosynthetic gene clusters in bacteria, Cell Chemical Biology (2021). DOI: 10.1016/j.chembiol.2021.08.009 Journal information: Cell Chemical Biology
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