Day 4: Massive genome engineering

Written by: Karmella Haynes.

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Harris Wang, tonight’s invited speaker, shared his lab’s work with MAGE, massive genetic engineering of bacterial chromosomes, in living cells. The new sequences are programmed into short bits of synthesized DNA, then fed into living bacteria, which incorporate these bits into their chromosome. This powerful technique allowed Wang to disrupt hundreds of genes and proteins, and to observe the effect of these changes on the fitness of bacteria. This massive project enabled greater insights into the function of bacteria’s molecular parts.

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David Savage, part of our course leadership team, placed Harris’ enabling technologies into the context of applications by presenting important outstanding questions in metabolic engineering, which is the practice of altering the genetic codes of cells (such as bacteria) to get them to produce high amounts of useful natural compounds. This is not a trivial goal because the dynamic conversion of one molecule into another occurs in a multifaceted, interconnected yarn-ball of pathways. This makes it difficult to choose which dial to turn and to predict what will happen as a result. Dave’s research team observed a peculiar and very useful property of a certain intermediate in pigment production. A partially constructed purple pigment glows fluorescent green, acting as a beacon to help track progress along a complex pathway. They applied MAGE technology to alter the conversion pathway, and the glowing beacon enabled them to track the results. This lead to his new system RApid Metabolic Bacterial Optimization, or RAMBO. He also aims to reconfigure one of the most popular glowing molecular beacons, green fluorescent protein, into a new type of beacon to guide engineers through the maze of biochemical pathways.

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