Northwestern University

Benjamin James Des Soye

Michael Christopher Jewett

Rey William Martin

Yong Chan Kwon

Michael Christoper Jewett

Seok Hoon Hong

Samuel Ryan Davidson

Disclosed are compositions, methods, and kits for performing cell-free RNA transcription and/or cell-free protein synthesis (CFPS). The disclosed compositions, methods, and kits include or utilize components prepared fromspecies such as cellular extracts from

Cell-free protein synthesis platform derived from cellular extracts of

Disclosed are methods, systems, components, and compositions related to genomically recoded and engineered organisms. The disclosed genomically recoded and engineered organisms may be used to prepare extracts for use in platforms and methods for preparing sequence defined biopolymers in vitro. In particular, the methods, systems, components, and compositions relate to genomically recoded and engineered organisms comprising a strain deficient in release factor 1 (RF-1) or a genetic homolog thereof, wherein the genomically recoded organisms have been engineered to express a heterologous RNA polymerase that may be utilized to express a target protein from a transcription template comprising a promoter for the heterologous RNA polymerase, such as bacteriophage T7 RNA polymerase.

Genomically recoded organisms lacking release factor 1 (RF1) and engineered to express a heterologous RNA polymerase

The invention relates to genomically recoded organisms, platforms for preparing sequence defined biopolymers in vitro comprising a cellular extract from a genomically recorded organism, and methods for preparing sequence defined biopolymers in vitro are described. In particular, the invention relates to genomically recoded organisms comprising a strain deficient in release factor 1 (RF-1) or a genetic homolog thereof and at least one of at least one additional genetic knock-out mutation, at least one additional upregulated gene product, or both at least one additional knock-out mutation and at least one additional upregulated gene product.

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Evanston, IL, United States of America

Benjamin James Des Soye