The Genome Engineering Center (GEC) at Washington University uses engineered endonucleases to create targeted modifications of mammalian genomes in order to speed scientific discovery, validate therapeutic targets, and increase scientific productivity.Through our cell engineering services, we work with investigators to create custom, user-defined cell lines. The GEC can generate modified cells with single base pair point mutations, small insertions or deletions, large insertions or deletions, gene knockout, and even multiplex modifications.We have experience using zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPRs) to create modified cells. The GEC creates novel, modified cell lines with the ultimate goal of advancing scientific discovery as a means to combat disease.http://genetics.wustl.edu/gec

Developed an iPSC model of familial platelet disorder with propensity to acute myeloid leukemia. Derived iPSCs from FPD patients and used the cells to model hematopoietic defect “in the dish”. Corrected RUNX1 mutation using zinc-finger nuclease mediated gene targeting at the RUNX1 locus, and showed hematopoietic defect was corrected. Successfully targeted a RUNX1-luciferase reporter cassette to the RUNX1 locus in a wildtype iPSC line. Used reporter cells to perform high-throughput small molecule screening and identified candidate drugs with the ability to induce RUNX1 expression.

Created a knock-in mouse model of a generic monogenic disease to serve as a paradigm for gene therapy. Studied zinc finger nuclease mediated gene targeting in a variety of primary cells isolated from the mouse and determined feasibility and frequency of correcting the mutant gene. Achieved gene targeting in murine embryonic stem cells and evaluated pluripotency of targeted cells. Researched toxicity of zinc finger nucleases in primary cells. Skills employed: Designing and cloning of transgenic constructs and expression vectors, maintenance of ES, primary, and transformed cell cultures, FACS, qRT-PCR using ABI Prism II, Southern blot analysis, westerns, IHC, fluorescent microscopy, teratoma formation assays, transfection techniques for cell cultures, and working with lentivirus (cloning, virus production, and titering). Initiated and maintained a transgenic mouse colony from 2004-2009. Gained experience in sequencing programs (Vector NTI, FinchTV), directed and supervised rotation students working on independent projects.

Evaluated incoming lots of kits for appropriate type and quantity of reagents then transferred to storage area. Assisted with inventory control.

Screened 25 Salmonella samples for 4 different integron sequences in order to determine the relative frequency that they may be found in the southwestern area of Texas. Skills employed: bacteria culture, DNA extraction, PCR, gel electrophoresis.

Processed transgenic corn samples by extracting total protein and determined percent protein of interest. Also prepared media and performed plated reading on a spectrophotomer. Skills employed: Bradford assays, ELISAs, western blots, keeping a lab notebook. Gained experience in the use of the Packard Multiprobe II.