In brief• Current therapies fail to effectively treat cancer patients, largely due to the
difficulty in distinguishing between healthy and cancer cells
• We designed a CRISPR-based synthetic system that will be assembled andactivated only in cancer cells
• Proof-of-concept results show that the Boomerang system is highlyselective and does not affect healthy cells
• The system can be easily adapted for every patient and tumor using theindividual tumor characteristics
Klac BD
Mutant20,000 people die
of cancer every day
Our system
Results
Fig. 2. eGFP and mCherry expression, under human TERT and survivin promoters,respectively, in fibrosarcoma cancer cells (upper panels) vs. healthy fibroblasts (lowerpanels), using calcium phosphate plasmid transfection. Results show specific expression incancer cells.
This collaboration can take the Boomerang system to the next level ofcancer detection&diagnostics. Selective expression of the Affibody incancer cells can synergistically increase both systems` potential.
Human Practices
Culture and educationSchool workshopEducational convention
• Increased accessibility toinformation about syntheticbiology by arranging uniqueevents
• Establish a dialogue with industryexperts to improve our project
• Successful collaborations withiGEM teams
• Collaboration and advice panelswith leading experts in cancerresearch, synthetic biology, andbioethics
Ethics and philosophyEthical aspects of CRISPR use
Arranged panelsExperts from the fields ofbasic and clinical cancerresearch and synthetic biology
Special meetingsMinistry of HealthDirectors of the Oncology Dept.at Soroka Medical Center and Sheba Cancer Research Center
Public outreachRadio , Video, Media, Social networks: FB, Twitter Thousands of followersExternal website Science on the Bar event Conventions
CollaborationChina-Amoy, articleFrance-Aix-Maseille, Israel-Technion, convention
EntrepreneurshipMeeting with synthetic biologycompaniesMeeting with Venture CapitalCrowd funding
Future vision
The third part of the Boomerang system is a construct that,upon activation, drives the expression of any gene of interestfor multiple applications:1. Diagnostics through a secretion of protein biomarker2. Visualization of the tumor for surgical removal3. Inducing apoptosis/cell death specifically in cancer cells
The Boomerang system will allow easy configuration of thethree-plasmid CRISPR-based gene activation to adapt thetreatment to every tumor and patient. This way side-effectscould be minimized. Moreover, constant adaptation of thedesigned constructs will ensure the effectiveness of the system,despite the tumor heterogeneity.
Cancer-specific expression of gRNA and Cas9 will activate a third constructcontaining a synthetic promoter that can drive the expression of any targetprotein
One Boomerang - Three Applications
Motivation
Fig. 1. hTERT and Survivin expression levels in several human cancer cell types, evaluated byqPCR. The results confirm the suitability of these two promoters for Boomerang system.
Fig. 3. Left panels: Similar AAV transductionefficiency in cancer and healthy cells. Rightpanels: Viral transduction of eGFP andmCherry under human TERT and survivinpromoters, showing specific expression incancer cells.
Collaboration
“Master” template – for modular cloning
Synthetic activation promoter
RGR design for gRNA – Ribozyme-gRNA-Ribozyme
Design HighlightsResearch Aim
phTERT
pSurvivin
Acknowledgments
Lab Workflow
Fig. 4. The activation of the entireBoomerang system. The three-plasmidCRISPR-driven eGFP expression is shownexclusively in cancer cells (upper panels), vs.no detected signal in health cells (lowerpanel).
• To design a modular system that willtransform the tumor into a in-cell drugfactory
• To experimentally validate the 3-component CRISPR-based Boomerang geneactivation system in human cancer cells
Stockholm team -Affibody approaches
Confocal imaging of Affibody expression in human fibrosarcoma – BGU team
