Harvard College

Wisconsin Alumni Research Foundation

Srivatsan Raman

George M Church

Noah D Taylor

Jameson K Rogers

Xiangyang Liu

Aseem Zoe Ansari

Jose Arcadio Rodriguez-Martinez

Phil Huss

A cyanuric acid-responsive biosensor includes a host strain or cell-free system including a first and a second expression cassette; the first expression cassette including, in operable communication, a constitutive promoter and a gene encoding an AtzR cyanuric acid-responsive transcription factor; and the second expression cassette including, in operable communication, a promoter regulated by the cyanuric acid-responsive transcription factor and a gene encoding a reporter protein, wherein the first expression cassette expresses the AtzR cyanuric acid-responsive transcription factor, and, in the presence of cyanuric acid, the AtzR cyanuric acid-responsive transcription factor drives expression of the reporter protein.

Sensor for cyanuric acid detection

Described herein is a method of preparing an unbiased library of phage variants, comprising (a) preparing a population of 'acceptor phage'; (b) removing an endogenous target gene and inserting gene variants into the acceptor phage genomes; (c) enriching the recombined phages; and (d) expressing the library for selection. The acceptor phage is a lytic phage comprising a synthetic genome wherein the target gene of interest is flanked by recombinase sites. The acceptor phage infects a first host bacteria expressing a recombination plasmid facilitating recombination. The phages then infect a second host bacteria expressing a counterselection system that accumulates recombined phage variants and selecting against non-recombined phages. The accumulated phage variants infect a third host bacteria. The phage library may then be sequenced and characterized.

Methods of making unbiased phage libraries

The present invention relates to genetically modified bacteria and methods of optimizing genetically modified bacteria for the production of a metabolite.

Methods for selecting microbes from a diverse genetically modified library to detect and optimize the production of metabolites

Methods and compositions for making and isolating allosteric DNA binding proteins that bind to one or more allosteric effectors to induce a conformation change in the proteins are provided.

De novo design of allosteric proteins

Described herein is a method for identifying synthetic inducible promoters that have specified induction and/or repression for DNA binding proteins such as an allosteric transcription factor and an inducer molecule. The method includes an in vitro selection from an unselected polynucleotide library comprising a plurality of random degeneracies, and an in vivo selection to produce an induced promoter library. Produced is an induction table, which allows the selection of a promoter with specific induction and/or repression properties. Also included are biosensors containing the synthetic inducible promoters.

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Arlington, MA, United States of America

Srivatsan Raman