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@alex_nguyen_ba and I designed an in vitro assay to measure the stability of Cre-Lox integration products and tested a large number of inverted repeat mutations from the literature.pic.twitter.com/JK0RBbUJaU
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If the product of recombination contains a doubly mutated Lox site, the reaction equilibrium shifts towards integration.pic.twitter.com/FthmNV8CDP
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An ingenious solution to this problem takes advantage of mutant Lox inverted repeats (IRs) thought to decrease Cre binding. Since Cre binding to Lox IRs is cooperative, a site with a single mutated IR will recombine normally but a double mutant site will not.pic.twitter.com/rYtWqqrqIz
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Our system requires that the barcodes we integrate into the genome stay there,even after hundreds of generations of evolution. Unfortunately, Cre-Lox integration is reversible. Worse yet, at equilibrium, integration substrates are favored over integrated product.pic.twitter.com/YMF4vPfrcG
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I wish graphviz could draw this prettypic.twitter.com/Fdbfr9Aq3s
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