Sintezna biologija: medicina

Medicinske aplikacije Zdravstvena industrija (healthcare industry) predstavlja eno najmočnejših panog in največjih svetovnih tržišč. Sintezna biologija za medicinske aplikacije deluje v smeri pridobivanja cenejših zdravil in izboljšanih materialov. Hkrati pa si prizadeva za razvoj boljših diagnostičnih sredstev, preprečevanje razvoja bolezni in zdravljenje bolezni in okvar. Področja delovanja: biosenzorji za prepoznavanje molekul (senzorske molekule ali mikroorganizmi), pametna in poosebljena zdravila, vektorski in dostavni sistemi za zdravljenje, izboljšane lastnosti celic, tkivno inženirstvo  regenerativna medicina. Synthetic circuit development for the treatment of disease. Synthetic gene networks are uploaded into cells to therapeutically target the body’s endogenous networks, causing a transition from disease to healthy state. Here, the uploaded network is a bistable toggle switch, which enables cellular memory with a network of two mutually repressible modules.

Two-pronged attack strategy for biofilm removal with enzymatically active DspB-expressing T7DspB phage. Initial infection of E. coli biofilm results in rapid multiplication of phage and expression of DspB. Both phage and DspB are released upon lysis, leading to subsequent infection as well as degradation of the crucial biofilm EPS component, β-1,6-N-acetyl-d-glucosamine Genomes of engineered phage used for biofilm treatment. (A) Genome of T7select415-1 shows a unique BclI site and capsid gene 10B. (B) DspB-expressing phage T7DspB was created by cloning T3 gene 1.2 into the unique BclI site and cloning the ϕ10-dspB construct after capsid gene 10B. (C) Non-DspB-expressing control phage T7control was created by cloning T3 gene 1.2 into the unique BclI site and cloning the control S·Tag insert (included in the T7select415-1 kit) as a fusion with the capsid gene 10B. PNAS 104 (27) 11197–11202, 2007

Assays for E. coli TG1 biofilm levels and phage counts after 24 h with no treatment or with treatment with phage T7wt, phage T3wt, non-DspB-expressing phage T7control, or DspB-expressing phage T7DspB. Error bars indicate SEM. (A) Mean absorbance (600 nm) for n = 16 biofilm pegs stained with 1% CV, solubilized in 33% acetic acid, and diluted 1:3 in 1× PBS (50). (B) Mean cell densities [log10(CFU per peg)] for n = 12 biofilm pegs. Pegs treated with T7DspB resulted in a 3.65 log10(CFU per peg) reduction in viable cells recovered from E. coli biofilm compared with untreated biofilm. (C) Mean phage counts [log10(PFU per peg)] recovered from media in n = 3 microtiter plate wells (wells) or sonication of n = 3 biofilm pegs (biofilm), as indicated, after 24 h of treatment with initial inoculations of 103 PFU per well. Both T7control and T7DspB showed evidence of replication with phage counts obtained from the microtiter plate wells or with phage counts recovered from the biofilms after sonication. PNAS 104 (27) 11197–11202, 2007

Synthetic biology approaches for treatment of bacterial infection and cancer. (A) Engineered bacteriophage boosted antibiotic killing efficacy by disrupting repair of antibiotic-induced damage. (B) Engineered bacteria invaded cancer cells and knocked down a cancer gene using RNAi. Science 333(6047) 1248-1252, 2011

Synthetic biology approach for microbiome engineering Synthetic biology approach for microbiome engineering. Native commensal bacteria were engineered to secrete the molecular signal cholera autoinducer (CAI-1), which leads to inhibition of V. cholerae virulence. Science 333(6047) 1248-1252, 2011

Diseases can be targeted with new synthetic biology methods for cell therapy and regenerative medicine. (A) Urate homeostasis was restored in vivo by the delivery of cells with a synthetic circuit. Uric acid induced the derepression of an engineered urate oxidase, which then lowered uric acid levels in mice. (B) Synthetic modified RNAs encoding the KMOS transcription factors were delivered to mammalian fibroblasts to induce pluripotency upon translation. The RNA-induced pluripotent stem cells could be driven down numerous cell lineages. Science 333(6047) 1248-1252, 2011

disseminated in a population. A synthetic gene drive for disrupting malaria vector capacity in mosquitos. In the model system, a transgenic mosquito carrying the gene drive (M1) mates with a transgenic mosquito containing endonuclease recognition sites within a reporter expression sequence (M2). Offspring of this pairing contain a chromosome from each parent. The endonuclease in the gene drive creates a double-stranded break in the opposite (reporter) chromosome and endogenous cellular processes use the chromosome containing the gene drive as a template for repair. As a result, both chromosomes in all offspring soon contain the gene drive and the interruption is quickly disseminated in a population. Science 333(6047) 1248-1252, 2011

template, and carries no risk of infection by attenuated pathogen. Synthetic biology approach for vaccine development. Immunostimulatory, antigenexpressing liposomes constructed with bacterial transcriptional and translational machinery produce model antigens in vivo for vaccination. In live mice, antigen-expressing liposomes generated a higher humoral immune response compared to control vaccines (liposomes encapsulating only the antigen, the transcription-and-control network, or the DNA template, respectively). This system can be easily altered for other antigens by simply changing the DNA template, and carries no risk of infection by attenuated pathogen. Science 333(6047) 1248-1252, 2011

regulate gene expression in mammalian cells. Controlling mammalian gene expression. A synthetic circuit containing an inducible repressor module and an RNA interference module was constructed to tightly and tunably regulate gene expression in mammalian cells. Science 333(6047) 1248-1252, 2011

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