Les outils de sélection : les nouveaux besoins industriels. Philippe Gabant, CSO

Transcription

1 Les outils de sélection : les nouveaux besoins industriels Philippe Gabant, CSO pgabant@syngulon.com

2 Applications of microorganisms in industries 2

3 Synthetic Biology Biofuels made from algae that will be able to scale, and compete with oil, will have to be synthesized and will not come from nature, says genomics scientist and entrepreneur Craig Venter. Oct. 23,

4 Vision of Syngulon The facts 1. Tools developed initially for sequencing human genome economically become accessible 2. Need for new technologies to meet the specific constraints in industrial biotechnology it may be possible to create high-value substances such as biofuels by modifying simple organisms such as E. coli bacteria. May 2013, Next-generation genomics in Disruptive technologies: Advances that will transform life, business, and the global economy (McKinsey Global Institute) 4

5 Synthetic Biology New applications of microbial in Bioindustry: Microrefineries and production in (semi-)open plants 5

6 The Problem How to control industrial KEY ISSUES Microorganisms? 1. Genetic security 2. Industrial Genetic Bio-control to increase yield 3. Protection of the micro-refineries against microbial invaders 4. Genetic containment 6

7 The Solution New selection technology designed for Industrial biotechnology Killing the industrial Microbes (micro-refineries) outside of the refineries: Security and Gene containment Control bacterial invaders: genetic shielding Increase production stability of the micro-refineries "Give me a fulcrum and I will move the world," from Archimedes, gets updated in the synthetic genomics lab to, "Give me a selection, and I will change the world." George M. Church in "Regenesis: How Synthetic Biology Will Reinvent Nature and Ourselves" 7

8 Technology Genetic selection (definition) A selection is an approach allowing only a sub-population of interest to survive The result is an enrichment of only individuals presenting the behavior (phenotype) of interest (ie production of a certain compound) 8

9 Toxin-Antitoxin Technologies Development of selections for genetic engineering At ULB in the 90 we designed selection based on bacterial suicidal poison-antidote. Natural bacterial loci: initially isolated from plasmids The paradigm: ccd locus of the F plasmid O/P ccda Antidote ccdb Poison Small genes; CcdB inhibits DNA gyrase and CcdA counteracts CcdB killing activity 9

10 Toxin-Antitoxin Technology CcdB CcdB CcdB CcdB Plasmid Plasmid CcdB CcdB Inside Outside 10

11 Toxin-Antitoxin Technology 1 Genetic engineering selection standard 11

12 Toxin-Antitoxin Technology 2 Plasmid Antidote (CcdA) Poison (CcdB) Target Gene of interest Need: replacement of antibiotic selection Approach: use of the bacterial poison gene (ccdb) and its natural antidote (ccda). During production bacteria loosing the plasmid are killed (suicide mechanism) Applications: antigen production, recombinant biocompounds production, pdna Closed and controlled batch production mono-cellular Key issues Selection of recombinants / expressing clones Alternative to antibiotic selection (to follow regulatory guidance) Minor issues Genetic containment (via physical containment, sterile environment) Non-issues: control of contaminants, partners control, security/competitors (ULB patents, Licensing by Delphi Genetics) Technology licensed by Delphi Genetics to: 12

13 Industrial biotechnology environment Inside Genosphere Outside 13

14 Need of a selection to design a Genetic Firewall Outside Inside 14

15 New Selection context = New Genes need to be used Micro-refineries are in interaction with the environment: Security and Gene containment issues Control bacterial invaders: genetic shielding Increase production stability of micro-refineries Development of a genetic firewall based on Bacteriocins Selection Loci Genosphere 15

16 New Selection Technology for industrial biotech Immunity Plasmid Bacteriocin (molecule naturaly secreted to kill or control other bacteria) Target Gene of interest Need: selection not only focused on the host of interest (suicidal approach) but also allowing the production microorganisms (micro-refineries) to control its growing environment and limit the spreading of micro-refineries outside the production plan Approach: bacterial bacteriocins loci are adapted for a mutidimensional selection controlling the hosts and environment during production Applications: genetic vigilance of the production plant allowing continuous production Open / semi-open continuous production could be multicellular Key issues Genetic containment (important bacteria express potential pollutants) Genetic security / competitors Selection of recombinants / expressing clones Alternative to antibiotic selection (to protect environment) Control of contaminants (bacterial invaders, divergent micro-refineries, escaping host) Partners control Non-issues (because impossible): physical containment, sterile (Syngulon patent pending, August 2013) 16

17 The Competition Indirect Competition: Non GMO bacteria used in existing processes Direct Competitors: Other selection technologies Selection technologies respective advantages Nat Nat+T ATB T-AT Security Gene containment Yield optimization Genetic shielding Open process ATB = antibiotics T-AT = toxin-antitoxin Nat = nature Nat+T = nature + constraint as T 17

18 Q & A Philippe Gabant, CSO pgabant@syngulon.com 18