Cell Engineering Shared Resource

Wake Forest University Health Sciences, Winston-Salem NC

Investigators

Linked publications, trials & patents

Abstract

CELL ENGINEERING SHARED RESOURCE (CESR): PROJECT SUMMARY The objective of the Cell Engineering Shared Resource (CESR) is to offer expertise in cell line engineering using viral and non-viral approaches and to provide the resources to analyze cancer cell growth/death in vitro and in vivo under a wide variety of experimental conditions. The use of various viral and non-viral vectors, as well as gene editing techniques (CRISPR/Cas9) have become critical in the experimental toolbox of cancer researchers to effectively manipulate gene expression and to create desired mutations. CESR has expertise in designing, implementing, and optimizing new cell culture, cell transduction, and cell line engineering procedures for investigators and can conduct short-term experiments in a cost-effective manner. CESR provides consultation without charge for project design. CESR offers ready-to-use stocks of lentiviruses for fluorescent and luminescent labeling and packages any retrovirus, lentivirus, or adeno-associated virus transfer plasmids into a replication-deficient viral vector. Additionally, CESR uses Gateway cloning to insert genes of interest into viral vector transfer plasmids, enabling generation of viral vectors from any cDNA. CESR is a member of Sigma- Aldrich’s CRISPR Core Partnership Program, which enables CESR users to directly work with scientists at Sigma-Aldrich to have sgRNAs designed to suit their needs. CESR stocks ready-to-use lentivirus-based Cas9 expression vectors, produces and delivers Cas9-gRNA ribonucleoproteins to cells for gene editing, and evaluates on-target cutting efficiency of CRISPR/Cas9 through quantification of in/del frequency using targeted PCR and DNA resolvase-based approaches. CESR establishes cell lines from primary patient samples and maintains a regional partnership with the Duke University Functional Genomics Facility for high -throughput screening of small molecules and CRISPR/shRNA libraries. CESR manages advanced instrumentation for monitoring cell growth in vivo and in vitro, including an IncuCyte Zoomfor live fluorescence imaging, a BioSpherix Xvivo X3 Hypoxia Chamber, a Nanosight NS500 for measurement of exosome/virus concentration and size, two IVIS Lumina III systems for in vivo fluorescence and luminescence imaging, and, most recently, an iThera MSOT inVision 256-TF for full body, multispectral optoacoustic imaging of rodents to study physiological processes at a molecular and cellular level. CESR provides access to over 200 characterized cancer cell lines, offers liquid nitrogen storage for members’ cell lines, conducts mycoplasma testing, procures nucleotide primers, offers training for instrumentation and advanced tissue culture/cell line transduction, and prepares custom media formulation. CESR also offers an on-site stockroom for tissue culture reagents, including media and sera, and custom procurement services with substantial discounts from multiple vendors. Over the project period, 106 WFBCCC members used CESR, representing 51% of total users – up from 44% at the time of the previous renewal. The number of peer-reviewed funded users also increased to 55 compared to 52 in the previous cycle. CESR provided support for 120 publications and 55 peer-reviewed, cancer-focused grants.