A new method to deal with multi-resistant pathogens by damaging resistance genes or the genome with CRISPR/Cas using bacteriophages transduction

Daniel Derndorfer (daniel.derndorfer@edu.uni-graz.at)

My idea is to create a method to damage the virulence genes within pathogens by using the CRISPR/Cas-method. Maybe it is possible to reduce or even stop the horizontal gene transfer of resistance genes within a patient and a subsequently therapy or antibiotic treatment is possible to defeat the infection. Using bacteriophages, which are highly specific to a bacterial host, the pathogen could be transduced with a CRISPR/Cas-plasmid, encoded with the sequence of a resistance gene as guide RNA. If the transduction is successful and the plasmid remains stable inside the cell, the bacterial transcription- and translation system produces a functional active CRISPR/Cas system. The guide RNA, connected with Cas, binds to the gene of interest, e.g. resistance genes, and lets the endonuclease damage (not simply cutting out) the gene. This should lead to sensitive cells and even if there would be a gene exchange there is no functional gene because of the damage. There is no actual data of this proceeding, but I want to develop that idea to a practical method.

I‘m also highly interested in an ethical debate about CRISPR. Since this is a new tool, we, the scientific community, should take care about the correct application and propose rational arrangements to politics. In my opinion, this is a rare chance to visualize genetic modification in a good light for the public population. A fundamental refusal, as we see in most of the genetic debates, would disappear if there are moral standards and clearly defined safety rules. That would make working in this field a lot easier for everyone. The scientific community should be aware of their responsibility of using a new and effective tool for the good, for knowledge and for solving problems instead of improper use for personal profits.

Dynamic Metabolic Engineering with CRISPR/cas9

Niklas Farke (niklasf@gmx.net) & Kai Libicher (KaiLibicher@gmx.de)

 Metabolic engineering aims at increasing the productivity of a microbe by means of manipulating native metabolic pathways or by genetically inserting new ones. A frequently encountered problem is that some modifications negatively affect cell growth. This problem renders a lot of metabolic engineering efforts useless to industrial applications because the right balance between productivity and yield needs to be established first. CRISPR/cas9 has the potential to improve the situation. The idea is to transform the (only slightly modified) microbe with an inducible CRISPR/cas module(on a plasmid). After a certain biomass is reached the module is activated. Subsequently cas9 will introduce specific knock-outs to redirect all ressources towards product formation thereby putting an halt to the growth rate. Thus production yield would be increased precisely when growth is no longer needed.

This method will dramatically expand the metabolic engineering toolbox which by now is limited to static improvements. As the changes are introduced during the process we term the concept “dynamic metabolic engineering”. Another possible application of this could lie in programming rational responses to stresses during fermentation in the bioreactor, for instance when temperature control falls out or during power shortages.

CRISPR and Microbiome-hacking 

Dr. Julian Chollet (julian@sonnenstreifen.de)

The human gut microbiome consists of about 10.000 different microorganisms and is highly important for health and disease. Probiotic bacteria are commonly used to improve general well-being and to restore normal digestion in patients with diarrhea, constipations as well as severe clinical conditions like inflammatory bowel disease. Unfortunately, it has been shown that most commercial probiotics are not effective. A more drastic measure for the restoration of the colonic microflora is a procedure known as fecal microbiota transplant (FMT), or stool transplant. Here a whole community of fecal bacteria is transplanted into the patient from a healthy donor.

Would it be possible to immunize probiotic bacteria? I would like to take a laboratory derived Bifidobacterium and genetically insert the CRISPR-spacers that it might need to survive in my gut. When consumed, these strains might have a higher chance of survival than the usual commercial probiotics. Actually this bacterium should not even be considered transgenic, as it may acquire the same kind of immunity in a natural way. Generally the described procedure should not pose any danger to human gut ecology as long as non-pathogenic bacteria are used. These strains are only modified in a way they could and would also be modified in their natural environment.

CRISPR-FEM – Reclaiming female reproductive technologies through vaginal flora modification

Mary Maggic (maggic@mit.edu)

There is a huge need for a non-hormonal contraception that circumvents the institutional and pharmaco-industrial complexes that are dictating women’s bodies. Combining the elements of kitchen laboratory, DIY feminist self care, and low-cost access to CRISPR technologies, we can create a disruptive technology and citizen alternative to the Pill. If 1950s housewives, confined to the kitchen under patriarchal standards had access to CRISPR, what would they do with it?

I am proposing to explore this “thought experiment” (series of sequential questions) of kitchen laboratory science for DIY feminist health care using CRISPR technologies:

  1. How feasible and actionable is CRISPR in a kitchen laboratory setting?
  2. What are the potential applications if CRISPR were to be used to modify some of the most intimate microbes: vaginal microflora?
  3. Could CRISPR modification of vaginal flora be used for feminist gains, such as eliminating the use of hormonal contraceptives, circumventing institutional control, and gaining greater gynecological “know-how” over one’s body?
  4. What are the practical risks of such a CRISPR technology? Is it safe? Is it reversible?
  5. What are the ethical dilemmas with such technology, and do the pros outweigh the cons?