Zymergen Inc.

Philip Weyman

Kedar Patel

Sara Da Luz Areosa Cleto

Aaron Miller

Shawn Szyjka

Stephen Blaskowski

Solomon Henry Stonebloom

Colin Scott Maxwell

Sharon Nademanee

Elizabeth Lauren Meier

Melissa Netwal

Cameron Coates

Erik Jedediah Dean

Sean Poust

Vytas SunSpiral

William Serber

Matthew Jonathan Myers

Kunal Mehta

The present invention relates to methods for editing the genome of a microbial host cell in successive rounds of transformation. The method allows the introduction of genetic edits into the genome of a microbial host cell in an iterative fashion that does not require the use of functional counterselection following at least one round of transformation. It can be used to rapidly stack genetic edits in the genome of a microbial host cell. Kits for performing the methods are also disclosed.

Iterative genome editing in microbes

Aspects of this disclosure relate to liquid-based workflows for strain engineering. In strain engineering, cells undergo a modification process to acquire a desired change or changes in the cells. Some of the cells accept the change or changes and some do not. Provided herein are liquid-based methods for selecting the population of cells that have accepted the change or changes. Also provided herein are liquid-based methods for removing markers from transformed cells and liquid-based methods for isolating clonally pure populations of cells.

Liquid based selection and cell isolation

The present disclosure relates to methods of joining three or more double-stranded (ds) or single-stranded (ss) DNA molecules of interest in vitro or in vivo. The method allows the joining of a large number of DNA fragments, in a deterministic fashion. It can be used to rapidly generate nucleic acid libraries that can be subsequently used in a variety of applications that include, for example, genome editing and pathway assembly. Kits for performing the method are also disclosed.

Multiplexed deterministic assembly of DNA libraries

The present invention relates to methods for editing the genome of a microbial host cell in one or more rounds of transformation. The method allows the introduction of genetic edits into the genome of a microbial host cell in a pooled and/or iterative fashion that does not require the use of functional counterselection following at least one round of transformation. It can be used to rapidly stack genetic edits in the genome of a microbial host cell. Compositions and kits for performing the methods are also disclosed.

Pooled genome editing in microbes

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Alameda, CA, United States of America

Philip Weyman