GSE228761 Genetic circuits encoded in the Escherichia coli Nissle genome, applied to a simplified design for a phenylketonuria (PKU) treatment

Contributors : Alexander J Triassi ; Brandon D Fields ; Catherine E Monahan ; Jillian M Means ; Yongjin Park ; Hamid Doosthosseini ; Jai P Padmakumar ; Vincent M Isabella ; Christopher A VoigtSeries Type : Expression profiling by high throughput sequencingOrganism : Escherichia coli Nissle 1917As living therapeutics, Escherichia coli Nissle 1917 (EcN) have been genetically engineered to express enzymes that degrade toxic metabolites, immunoregulators to fight cancer or antimicrobial molecules to kill pathogens. Carrying these functions can impose a burden if constitutively expressed and, in the body, plasmid stability cannot be enforced with a selectable marker. Carrying these functions in the genome has the potential to improve stability, but expression is lower and genetic insertions can interfere with native genes. Here, we insert and optimize multiple insulated landing pads in the EcN genome and develop a protocol to accelerate integration. Synthetic regulation, including seven inducible systems and seven repressor-based NOT gates, are characterized in the genome, showing up to 130-fold induction. These tools were applied to engineer a strain undergoing clinical trials as a treatment for phenylketonuria (PKU) (EcN SYNB1618). We moved the pathway for phenylalanine to trans-cinnamic acid (TCA) conversion (pheP, stlA) to a genomic landing pad under synthetic regulatory control, including an inverted inducible system to simplify manufacturing. The resulting strain achieved ...
Source: GEO: Gene Expression Omnibus - Category: Genetics & Stem Cells Tags: Expression profiling by high throughput sequencing Escherichia coli Nissle 1917 Source Type: research