Leah A. Santat

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The Notch-Delta signalling pathway allows communication between neighbouring cells during development. It has a critical role in the formation of 'fine-grained' patterns, generating distinct cell fates among groups of initially equivalent neighbouring cells and sharply delineating neighbouring regions in developing tissues. The Delta ligand has been shown(More)
RNAi is proving to be a powerful experimental tool for the functional annotation of mammalian genomes. The full potential of this technology will be realized through development of approaches permitting regulated manipulation of endogenous gene expression with coordinated reexpression of exogenous transgenes. We describe the development of a lentiviral(More)
Effective and stable knockdown of multiple gene targets by RNA interference is often necessary to overcome isoform redundancy, but it remains a technical challenge when working with intractable cell systems. We have developed a flexible platform using RNA polymerase II promoter-driven expression of microRNA-like short hairpin RNAs which permits robust(More)
Robert A. Rebres, Tamara I. A. Roach, Iain D. C. Fraser, Finly Philip, Christina Moon, Keng-Mean Lin, Jamie Liu, Leah Santat, Lucas Cheadle, Elliott M. Ross, Melvin I. Simon, and William E. Seaman 4 From the Alliance for Cellular Signaling, Northern California Institute for Research and Education and Department of Medicine, University of California,(More)
Cross-talk between Gα(i)- and Gα(q)-linked G-protein-coupled receptors yields synergistic Ca(2+) responses in a variety of cell types. Prior studies have shown that synergistic Ca(2+) responses from macrophage G-protein-coupled receptors are primarily dependent on phospholipase Cβ3 (PLCβ3), with a possible contribution of PLCβ2, whereas signaling through(More)
RNA interference (RNAi) was investigated with the aim of achieving gene silencing with diverse RNAi platforms that include small interfering RNA (siRNA), short hairpin RNA (shRNA) and antisense oligonucleotides (ASO). Different versions of each system were used to silence the expression of specific subunits of the heterotrimeric signal transducing(More)
A recombinant AAV2 (rAAV2) vector encoding antisense RNA to HIV-1 transactivating region (TAR) was evaluated for transduction of human cord blood CD34+CD38- hematopoietic stem cells (HSC) capable of serial engraftment in nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mice. Results revealed long-term multilineage marking in primary and(More)
Construct Name Promoter Gene Mammalian Selection Role in this work pEV‐UAS‐H2B‐citrine UAS H2B‐citrine Zeocin Reporter for hN1‐Gal4 pEV‐12xCSL‐H2B‐ citrine 12xCSL H2B‐citrine Zeocin Reporter for hN1 pcDNA3‐hN1‐mod1 CMV hNotch1 Neomycin hN1 construct pcDNA3‐hN1‐ mcherry CMV hN1‐mcherry Neomycin hN1 construct (used) pCDNA3‐hNECD – Gal4 CMV hNECD – Gal4(More)
The use of RNA interference to knock down protein phosphatases has proven to be a valuable approach to understanding the functions of these enzymes in mammalian cells. Many protein phosphatases exist as multisubunit and multigene families, which has made it difficult to assess their physiological functions using traditional approaches. The ability to(More)
Cellular responses to inputs that vary both temporally and spatially are determined by complex relationships between the components of cell signaling networks. Analysis of these relationships requires access to a wide range of experimental reagents and techniques, including the ability to express the protein components of the model cells in a variety of(More)