Gene Editing Shared Resource

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Please remember to cite the Shared Resources!

Research reported in this publication was supported by The Ohio State University Comprehensive Cancer Center and the National Institutes of Health under grant number P30 CA016058.

We thank the XX Shared Resource at The Ohio State University Comprehensive Cancer Center, Columbus, OH for (XX)

About Us (back to top)

Gene editing using CRISPR/Cas technologies is one of the most advanced, cutting-edge approaches to study gene, RNA and protein functions in a near-physiological setting by targeting the endogenous genetic code of the cells of interest. Furthermore, it can be used to design and validate novel diagnostic and therapeutic tools for a wide range of human diseases, including cancer.

The GEdSR is a crucial shared resource supported by the OSUCCC with the overarching mission to support the investigators of the OSUCCC for all their projects requiring genetic manipulations of established and primary cell lines using CRISPR-based approaches.

GEdSR can perform DNA-based (using recombinant plasmids) and DNA-free (using synthetic Cas9/RNP complexes) gene editing projects. In addition, GEdSR is provided with fully equipped lab space for the preparation of reagents required for gene editing projects, as well as tissue culture rooms for the generation of cell lines required by the investigators.

994 Biomedical Research Tower
460 West 12th Avenue
Columbus, OH 43210

Phone: 614-3662551

dario.palmieri@osumc.edu

Meet the Team(back to top)

Dario Palmieri, PhD - Director dario.palmieri@osumc.edu

Dr. Palmieri is an assistant professor in the Department of Cancer Biology and Genetics at The Ohio State University College of Medicine, where he studies how cancer cells dysregulate cell cycle progression and DNA damage response. He is also a member of the Cancer Biology Program and serve as director of the Gene Editing Shared Resource at the OSUCCC – James. Dr. Palmieri's research is focused on RNA and RNA-binding proteins, including their role in tumorigenesis and intercellular communication. The main goal is to gain insights on their biology, which is instrumental in developing new therapeutic approaches. His lab also has considerable experience in the use of CRISPR/Cas9 as a tool in editing endogenous genes to study their functions. Additionally, Dr. Palmieri serves as co-PI on a study funded by a Pelotonia Junior Investigator Award that examines a new mouse model that can be used to assess rigor and reproducibility of extracellular vesicles as cancer biomarkers. Dr. Palmieri has authored or co-authored several publications in such peer-reviewed journals as Journal of Experimental Medicine, PNAS and Oncogene. He is also a member of the American Society for Cancer Research and the International Society for Extracellular Vesicles.

Technical Director: Lara Rizzotto, PhD

Research Associate: Saranya Lamba

Scientific Advisor: Richard Fishel, PhD

Available Services (back to top)

The GEdSR provides tailored services using CRIPSR-based technologies to support the needs of all its users. Services include (but are not limited to):

Gene-editing services using immortalized cell lines and primary cells
Custom design of CRISPR knock-out and knock-in approaches
gRNA cloning and/or delivery as RNP complexes
Design of repair templates for small knock-ins and point mutations
Generation of vectors and cell lines expressing biosensors for CRISPR-based screens
Design and generation of large repair templates
Generation of CRISPR-KO/KI bulk cell lines or single clones
Cell sorting and live-cell imaging of gene-edited cells
Cas13-based RNA targeting
Approaches for gene editing without DNA double-strand breaks (base editing and PRIME edit)
Design and generation of libraries for CRISPR screenings

The GEdSR staff works with the investigators in a collaborative fashion thoroughly participating in the 1) study design; 2) identification of optimal strategies; 3) generation of preliminary data to assess project feasibility; and 4) generation of gene-edited primary and established cell lines.

Major equipment

Resources dedicated to the GEdSR include:

Dedicated lab space to produce reagents required for gene-editing projects (e.g., molecular cloning and cell genotyping)
Dedicated tissue culture room, including all the standard equipment required for manipulation of primary and established cell lines (2 biosafety cabinets, 2 centrifuges and 1 light microscope)
Lonza 4D Core Unit system equipped for optimal high-throughput electroporation (Lonza 96-well Shuttle)
EVOS M5000 microscope equipped with multiple light cubes for bright-field and fluorescence microscopy, magnification objectives ranging from 4X to 100X and onstage incubator for live-cell imaging
CountessFL automated cell counter
Sony MA900 cell sorter, equipped with four lasers and 12 detectors, capable of simultaneous sorting of up to four individual cell populations and single-cell plating. The chip-based system allows the selection of appropriate pressure conditions to maximize purity, yield and cell viability in the sorting process.

Click here for full list of services and fees

Latest News (back to top)

https://www.youtube.com/watch?v=Z1Oih40GlSo

We are planning to add the following services to our portfolio:

Gene editing of Pluripotent Stem Cells
Performance of pooled CRISPR screens, which provide the investigators with the unique possibility to simultaneously assess the importance of multiple genes on specific biological functions and pathways