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New cell programming method rapidly turns bacteria into biotech factories
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Harvard Medical School

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New cell programming method rapidly turns bacteria into biotech factories

Harvard Medical School researchers in the U.S. have developed a new cell programming method, called Multiplex Automated Genome Engineering (MAGE), which promises to give biotechnology, in particular synthetic biology, a powerful boost.

London, July 27 : Harvard Medical School researchers in the U.S. have developed a new cell programming method, called Multiplex Automated Genome Engineering (MAGE), which promises to give biotechnology, in particular synthetic biology, a powerful boost.

This work was carried out in the lab of George Church, a professor of Genetics, by a pair of researchers.

With the novel technique, the research team could rapidly refine the design of a bacterium by editing multiple genes in parallel instead of targeting one gene at a time.

They transformed self-serving E. coli cells into efficient factories that produce a desired compound, accomplishing in just three days a feat that would take most biotech companies months or years.

"We initiated the project to close the gap between DNA sequencing technology and cell programming technology," Nature magazine quoted graduate student Harris Wang, the paper's co-first author, as saying.

"The goal was to use information gleaned from genetics and genomics to rapidly engineer new functions and improve existing functions in cells. We wanted to develop a new tool and demonstrate how to apply it; we were determined to hand labs a hammer and a nail," added postdoctoral researcher Farren Isaacs, the other first author.

During the study, the researchers selected a harmless strain of the intestinal nemesis E. coli, and added a few genes to its solitary circular chromosome, coaxing the organism to produce lycopene, a powerful antioxidant that occurs naturally in tomatoes and other vegetables. They could then focus on tweaking the cells to increase the yield of this compound.

Traditionally, scientists would accomplish this type of transformation by using recombinant DNA technology, also known as gene cloning, a complicated technique that involves isolating, breaking up, reassembling, and then reinserting genes.

However, the researchers engaged in the current study took a different approach, blending an engineer's logic with a biologist's appreciation for complexity.

"Genes function in teams, not in isolation. Cloning often encourages us to ignore the interdependence of genes and oversimplify the cellular system. We might forget, for example, that one mutation can strengthen or weaken the effects of another mutation," says Wang.

"It's nearly impossible to predict which combinations of mutations will confer the desired behavior. Biology is so complex that we don't know the optimal solution," adds Isaacs.

Considering that, the researchers retooled evolution to generate genetic diversity at an unprecedented rate, increasing the odds of finding cells with desirable properties.

The E. coli bacterium contains approximately 4,500 genes. The team focused on 24 of these-honing a pathway with tremendous potential-to increase production of the antioxidant, optimizing the sequences simultaneously.

They divided the 24 DNA sequences up into manageable 90-letter segments, and modified each, generating a suite of genetic variants.

Then, armed with specific sequences, the researchers enlisted a company to manufacture thousands of unique constructs. The team was then able to insert these new genetic constructs back into the cells, allowing the natural cellular machinery to absorb this revised genetic material.

While some bacteria ended up with one construct, some ended up with multiple constructs.

According to the researchers, the resulting pool contained an assortment of cells, some better at producing lycopene than others.

Extracting the best producers from the pool, the team repeated the process over and over to further hone the manufacturing machinery. To make things easier, they automated all of these steps.

"We accelerated evolution, generating as many as 15 billion genetic variants in three days and increasing the yield of lycopene by 500 percent. Can you imagine how long it would take to generate 15 billion genetic variants with traditional cloning techniques? It would take years," Harris says.

The pathway the team refined plays a role in the synthesis of many valuable compounds, ranging from hormones to antibiotics, so the reprogrammed bacteria can be used for a variety of purposes. In addition, the MAGE platform itself unlocks new possibilities.

"We decided to engineer in the context of biology, embracing evolution rather than trying to fit a square peg in a round hole. This automated, multiplex technology will allow labs to engineer entire pathways and genomes and take cell programming to a whole new level," says Church.


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