BAR-ILAN INSTITUTE OF NANOTECHNOLOGY & ADVANCED MATERIALS | 2019 ANNUAL REPORT

CRISPR technology to treat malignancies using cancer-immunotherapy. Our strategy seeks to improve cancer-fighting potency of human T lymphocytes by genetic modification of immunoreceptors which play a key role during cancer immunity cycle. Publications 2018 and 2019 • A Jacobi, G Rettig, M Schubert, J Shapiro, O Iancu, A Tovin, E Cedrone, M Dobrovolskaia, A Hendel, M Behlke, “A comprehensive study comparing on-and off-target levels of the most common forms of CRISPR/ Cas9 guide RNAs”, European J of Human Gentics 27, 788-789, 2019 . • A Lev, AJ Simon, O Barel, E Eyal, E Glick-Saar, O Nayshool, O Birk, T Stauber, A Hochberg, A Broides, S Almashanu, A Hendel, Y N Lee, R Somech, “Reduced function and diversity of T cell repertoire and distinct clinical course in patients with IL7RA mutation”, Frontiers in immunology 10, 1672, 2019 . • MK Cromer, S Vaidyanathan, DE Ryan, B Curry, AB Lucas, J Camarena, Milan Kaushik, Sarah R Hay, Renata M Martin, Israel Steinfeld, Rasmus O Bak, Daniel P Dever, Ayal Hendel, Laurakay Bruhn, Matthew H Porteus, “Global Transcriptional Response to CRISPR/Cas9-AAV6-Based Genome Editing in CD34+ Hematopoietic Stem and Progenitor Cells”, Molecular Therapy 26 (10), 2431-2442, 2018 . • S Vaidyanathan, KT Azizian, AKMA Haque, JM Henderson, A Hendel, Sabrina Shore, Justin S Antony, Richard I Hogrefe, Michael SD Kormann, Matthew H Porteus, Anton P McCaffrey, “Uridine Depletion and Chemical Modification Increase Cas9 mRNA Activity and Reduce Immunogenicity without HPLC Purification”, Molecular Therapy-Nucleic Acids 12, 530-542, 2018 . • A Hendel, J Shapiro, O Iancu, K Mizrahi, A Tovin, A Jacobi, G Rettig, Mollie Schubert, Mark Behlke, “CRISPR Gene Correction for Severe Combined Immunodeficiency Caused by Mutations in Recombination- 67 Publications 2018 and 2019 • Gonen N, Lovell-Badge R., “The regulation of Sox9 expression in the gonad”, Curr Top Dev Biol.;134:223-252, 2019 . • Garcia-Moreno SA, Futtner CR, Salamone IM, Gonen N, Lovell-Badge R, Maatouk DM., “Gonadal supporting cells acquire sex-specific chromatin landscapes during mammalian sex determination.”, Dev Biol. 15;446(2):168-179, 2019 . • Gonen N, Meller A, Sabath N, Shalgi R., “Amino Acid Biosynthesis Regulation during Endoplasmic Reticulum Stress Is Coupled to Protein Expression Demands.”, iScience. 27;19:204-213, 2019 . • Styr B, Gonen N, Zarhin D, Ruggiero A, Atsmon R, Gazit N, Braun G, Frere S, Vertkin I, Shapira I, Harel M, Heim LR, Katsenelson M, Rechnitz O, Fadila S, Derdikman D, Rubinstein M, Geiger T, Ruppin E, Slutsky I., “Mitochondrial Regulation of the Hippocampal Firing Rate Set Point and Seizure Susceptibility”, Neuron. 5;102(5):1009-1024.e8, 2019 . • Gonen N, Sabath N, Burge CB, Shalgi R., “Widespread PERK-dependent repression of ER targets in response to ER stress”, Sci Rep. Mar 13;9(1):4330, 2019 . • Mevorach Zussman N, Gonen N, Kovo M, Miremberg H, Bar J, Condrea A, Ginath S, “Protracted postpartum urinary retention-a long-term problem or a transient condition?”, Int Urogynecol J. 2019 . • Gonen N., Futtner CR., Wood S., Garcia- Moreno AS., Salamone IM., Samson SC., Sekido R., Poulat F., Maatouk DM., Lovell- Badge R., “Sex Reversal Following Deletion of a Single Distal Enhancer of Sox9”. Science; 360 (6396):1469-1473, 2018 . • Hass J, Bassil R, Gonen N, Meriano J, Jurisicova A, Casper RF., “The VEGF and PEDF levels in the follicular fluid of patients co- treated with LETROZOLE and gonadotropins during the stimulation cycle”, Reprod Biol Endocrinol. 29;16(1):54, 2018 . Dr. Ayal Hendel The Mina & Everard Goodman Faculty of Life Sciences Member of BINA, Nano-Medicine Center Research Areas • Biotechnology • Genetic therapy • Genetic engineering • Developing CRISPR technology as a method of gene therapy for genetic diseases Abstract Precise and efficient CRISPR genome editing as a curative therapy for genetic disorders. We are in the midst of a revolution in genome editing and CRISPR-Cas9 technology was the spark. With unprecedented rapidity, this technology has provided a straightforward, robust, and specific method for genome editing. Our research focuses on developing genome editing as curative therapy for genetic diseases and cancer. Our lab is particularly interested in applying genome editing for gene therapy of hematopoietic genetic disorders such as severe combined immunodeficiency (SCID). SCIDs are a set of life threatening genetic diseases in which patients are born with mutations in single genes, and are unable to develop functional immune system. While allogeneic bone marrow transplantation can be curative for these disorders, there remain significant limitations to this approach. We believe that the ultimate cure for these diseases will be transplantation of gene-corrected autologous CD34+ hematopoietic stem and progenitor cells (HSPCs). To be able to apply this approach in the clinic, we must assure that the genome-editing technology is efficient and safe. Hence, our research focuses on developing an optimized CRISPR- genome editing for robust, locus- specific and non-toxic functional gene correction in HSPCs. Additional aspect of our research focuses on applying the