The mission of the Cell Engineering Shared Resource (CESR) is to serve the scientific needs of Wake Forest Baptist Comprehensive Cancer Center (WFBCCC) researchers by offering expertise in cell culture/cell line engineering/viral vector propagation, and providing the resources to analyze cell growth/death in vitro and in vivo under a wide variety of experimental conditions.
The Cell Engineering Shared Resource is capable of designing, implementing and optimizing new cell culture/transduction procedures on behalf of investigators, and can conduct short-term experiments efficiently in a cost-effective manner.
The CESR houses advanced instrumentation for monitoring cell growth in vivo and in vitro, offers an on-site stockroom and acts as a central site for purchasing and distributing specific culture media/reagents, molecular biology kits, and standard laboratory supplies.
The Cell Engineering Shared Resource is capable of designing, implementing and optimizing new cell culture/transduction procedures on behalf of investigators, and can conduct short-term experiments efficiently in a cost-effective manner.
The CESR houses advanced instrumentation for monitoring cell growth in vivo and in vitro, offers an on-site stockroom and acts as a central site for purchasing and distributing specific culture media/reagents, molecular biology kits, and standard laboratory supplies.
William Gmeiner, Ph.D., MBA. R21CA218933: preparation and concentration of lentivirus expressing GFP/Luciferase; transduction and selection of colorectal cancer cell lines.
Pierre Alexandre Vidi, Ph.D., R00CA163957: preparation, concentration, and titering of retrovirus expressing a DNA damage reporter gene (53BP1-mCherry).
Tim Pardee, M.D., Ph.D., R01CA197991: packaging CRISPR library in lentivirus; increasing scale of preparation to enable full coverage of target cells with 300-fold representation of all gRNAs according to methods described in Joung et al. Genome scale CRISPR-Cas9 knockout and transcriptional activation screening. Nature Protocols. 2017;12(4).
Zach McIver, M.D.: Large scale prep of AAV5 expressing DREADD (3.5 ml; 3.75 x 1011 GC/ml).
Bethany Kerr, Ph.D., R00CA175291: Transduction and selection of prostate cancer cells using lentiviruses expressing hCD117-GFP, hCD117-FFLuc, and control vectors. Cell lines based upon clonal selection and stable pools were generated.
George Kulik, Ph.D., R21CA182248: preparation, concentration, and titering of lentiviruses expressing MCL1 shRNA and control vector.
Ravi Singh, Ph.D., R01CA207222: preparation, concentration, and tittering of lentiviruses expressing ZEB1/shZEB1, NOX4, EGFP, or mCherry; transduction of breast and lung cancer cell lines.
Gagan Deep, Ph.D., R21CA199628: preparation, concentration, and tittering of retrovirus HIF2a and control vector, and lentivirus expressing SV40T antigen/mCherry; developed reverse transduction method to increase infection efficiency and selection of prostate cancer cell lines.
Bethany Kerr, Ph.D. R00CA175291: Gateway cloning of estrogen receptor-GFP fusion protein and lentivirus packaging
William Gmeiner Ph.D., MBA (CGM) R21CA218933: Design of CRISPR gRNA targeting DPYD.
K.C. Balaji, M.D.: Design of CRISPR gRNAs targeting MYC and MAX; plasmid preparation for transient transduction of target cells with CAS9/tracrRNA/gRNA plasmid; design of primers and evaluation of on-target cutting efficiency using Guide-it Mutation Detection Kit.
Ravi Singh, Ph.D., R01CA207222: Design of CRISPRs gRNA targeting ESRP1; plasmid preparation for transient transduction of target cells with CAS9/tracrRNA/gRNA plasmid; design of primers and evaluation of on-target cutting efficiency using Guide-it Mutation Detection Kit.
Pierre Alexandre Vidi, Ph.D., R00CA163957: preparation, concentration, and titering of retrovirus expressing a DNA damage reporter gene (53BP1-mCherry).
Tim Pardee, M.D., Ph.D., R01CA197991: packaging CRISPR library in lentivirus; increasing scale of preparation to enable full coverage of target cells with 300-fold representation of all gRNAs according to methods described in Joung et al. Genome scale CRISPR-Cas9 knockout and transcriptional activation screening. Nature Protocols. 2017;12(4).
Zach McIver, M.D.: Large scale prep of AAV5 expressing DREADD (3.5 ml; 3.75 x 1011 GC/ml).
Bethany Kerr, Ph.D., R00CA175291: Transduction and selection of prostate cancer cells using lentiviruses expressing hCD117-GFP, hCD117-FFLuc, and control vectors. Cell lines based upon clonal selection and stable pools were generated.
George Kulik, Ph.D., R21CA182248: preparation, concentration, and titering of lentiviruses expressing MCL1 shRNA and control vector.
Ravi Singh, Ph.D., R01CA207222: preparation, concentration, and tittering of lentiviruses expressing ZEB1/shZEB1, NOX4, EGFP, or mCherry; transduction of breast and lung cancer cell lines.
Gagan Deep, Ph.D., R21CA199628: preparation, concentration, and tittering of retrovirus HIF2a and control vector, and lentivirus expressing SV40T antigen/mCherry; developed reverse transduction method to increase infection efficiency and selection of prostate cancer cell lines.
Bethany Kerr, Ph.D. R00CA175291: Gateway cloning of estrogen receptor-GFP fusion protein and lentivirus packaging
William Gmeiner Ph.D., MBA (CGM) R21CA218933: Design of CRISPR gRNA targeting DPYD.
K.C. Balaji, M.D.: Design of CRISPR gRNAs targeting MYC and MAX; plasmid preparation for transient transduction of target cells with CAS9/tracrRNA/gRNA plasmid; design of primers and evaluation of on-target cutting efficiency using Guide-it Mutation Detection Kit.
Ravi Singh, Ph.D., R01CA207222: Design of CRISPRs gRNA targeting ESRP1; plasmid preparation for transient transduction of target cells with CAS9/tracrRNA/gRNA plasmid; design of primers and evaluation of on-target cutting efficiency using Guide-it Mutation Detection Kit.