Methods for control of flux in metabolic pathways
AssigneeGreenLight Biosciences, Inc.
InventorDaniel Klein-Marcuschamer
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Abstract
The invention pertains to a method for preparing cells that can be used as biocatalysts by inducing in them a growth-decoupled state, in which interferase inhibits the expression of genes except the ones that code for the pathway enzymes of interest. mRNAs that code for interferase-resistant products are overexpressed in the background of a metabolically-frozen cell. Enzymes that compete for a substrate or product of the pathway of interest may be altered such that the enzyme is sensitive to a site-specific protease, which protease is inducible in the host cell.
Claims20
A cell genetically manipulated to comprise:
- an interferase coding sequence, which interferase cleaves mRNA at a specific sequence motif;
- coding sequences for at least two enzymes involved in a pathway of interest, wherein the coding sequences are modified to be free of motifs cleaved by said interferase.
A cell genetically manipulated to comprise:
- a protease coding sequence operably linked to an inducible promoter, which protease cleaves proteins at a specific sequence motif;
- a coding sequence for at least one competing enzyme involved in a pathway of interest, wherein the coding sequence is modified to encode a motif cleaved by said protease.
Dependencies are inferred from the claim text.