82 understand, at the molecular level, how sex is determined during embryonic development, i.e., how does an embryo develop into either male or female. In mammals, sex determination is genetically driven with XY individuals developing as males and XX individuals developing as females. However, if the process of sex determination is impaired, patients appear as XY females or XX males (sex reverse). These are all classified as patients with Disorders of Sex Development (DSD), with a prevalence of 1: 2500-4000 newborns. In the lab, we use the mouse as a model system and are interested at understanding the regulation of the key transcription factors controlling gonad formation and how these interact with each other. We also study the role of the non-coding genome in mediating the process of sex determination and DSD pathologies. We employ cutting-edge techniques to address these questions including CRISPR/Cas9 genome editing, transgenic mice production, advanced sequencing techniques as well as microscopy and molecular biology. Developing an in vitro model to study the gonads To develop an in vitro system that allows to model the mammalian gonads, we use mouse Embryonic Stem Cells (ESC) and develop differentiation protocols towards gonadal cell type of the gonads. In addition, we use similar protocols with Human ESC and induced Pluripotent Stem Cells to model DSD patients in vitro. Furthermore, we perform tissue engineering to model the testis and the spermatogenesis process using these stem cell-derived somatic cells along with germ cells in a 3D culture system. Publications 2020 and 2021 • Eozenou C. *, Gonen N. *, Touzon MS. *, Jorgensen A., Yatsenko SA., et al., McElreavey K., Belgorosky A., LovellBadge R., Rajkovic A., Bashamboo A. “Testis formation in XX individuals resulting from novel pathogenic variants in Wilms’ tumor 1 (WT1) gene”. Proc Natl Acad Sci U S A, 117 (24): 13680-13688. PMID: 32493750, 2020. Dr. Hendel Ayal 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, locusspecific and non-toxic functional gene correction in HSPCs. Additional aspect of our research focuses on applying the 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 2020 and 2021 • Atar Lev, Yu Nee Lee, Guangping Sun, Enas Hallumi, Amos J Simon, Keren S Zrihen, Shiran Levy, Tal Beit Halevi, Maria Papazian, Neta Shwartz, Ido Somekh, Sarina Levy-Mendelovich, Baruch Wolach, Ronit Gavrieli, Helly Vernitsky, Ortal Barel, Elisheva Javasky, Tali Stauber, Chi A Ma, Yuan Zhang, Ninette Amariglio, Gideon Rechavi, Ayal Hendel, Deborah Yablonski, Joshua D Milner, Raz Somech. “Inherited SLP76 deficiency in humans causes severe combined immunodeficiency neutrophil and platelet defects”. Journal of Experimental Medicine, 2021. • Ido Amit, Ortal Iancu, Alona LevyJurgenson, Gavin Kurgan, Matthew S McNeill, Garrett R Rettig, Daniel Allen, Dor Breier, Nimrod Ben Haim, Yu Wang, Leon Anavy, Ayal Hendel, Zohar Yakhini. “CRISPECTOR provides accurate estimation of genome editing translocation and off-target activity from comparative NGS data”. Nature communications, 2021. • Ido Somekh, Atar Lev, Ortal Barel, Yu Nee Lee, Ayal Hendel, Amos J Simon, Raz Somech. “Exploring genetic defects in adults who were clinically diagnosed as severe combined immune deficiency during infancy”. Immunologic Research, 2021. • Daniel Allen, Michael Rosenberg, Ayal Hendel. “Using synthetically engineered guide RNAs to enhance CRISPR genome editing systems in mammalian cells”. Frontiers in Genome Editing, 2021. • Jenny Shapiro, Adi Tovin, Ortal Iancu, Daniel Allen, Ayal Hendel. “Chemical modification of guide RNAs for improved CRISPR activity in CD34+ human hematopoietic stem and progenitor cells”. CRISPR Guide RNA Design, 37-48, 2021. • Schaffer AA, Kopel E, Hendel A, Picardi E, Levanon EY, Eisenberg E. “The cell line A-to-I RNA editing catalogue”. Nucleic Acids Research. 48(11):5849-5858, 2020. • Shapiro J, Iancu O, Jacobi AM, McNeill MS, Turk R, Rettig GR, Amit I, TovinRecht A, Yakhini Z, Behlke MA, Hendel A. “Increasing CRISPR Efficiency and Measuring Its Specificity in HSPCs Using a Clinically Relevant System”. Molecular Therapy - Methods & Clinical Development, 2020.
RkJQdWJsaXNoZXIy NDU2MA==