CHEMICAL EMBRYOLOGY: TECHNOLOGIES FOR MANIPULATING AND VISUALIZING DEVELOPMENT Abstract The emergence of pattern during embryogenesis requires dynamic control of gene function with spatiotemporal precision. Genetic screens and transgenic models have revealed many of the molecular mechanisms that regulate this transformation, and the completion of multiple genome sequencing projects has provided a comprehensive list of developmental genes. Our efforts to understand embryonic patterning at the molecular and systems levels, however, have been limited by current technologies for studying embryological processes. New methods for controlling and visualizing gene function in vivo are needed. In this application we describe synthetic probes that will enable the detection of endogenous RNAs or tagged proteins in live organisms, with at least picomolar sensitivity. In particular, we will develop reagents that couple gene expression with lanthanide luminescence and utilize them to interrogate the molecular mechanisms of zebrafish embryogenesis. The proposed reagents build upon our expertise in synthetic chemistry and zebrafish embryology, and they complement reverse-genetic technologies previously developed by our laboratory. Using these chemical tools, developmental biologists will be able to simultaneously observe gene function and morphogenesis in real time, providing an unprecedented mechanistic view of embryogenesis.
| Payumo, Alexander Y; McQuade, Lindsey E; Walker, Whitney J et al. (2016) Tbx16 regulates hox gene activation in mesodermal progenitor cells. Nat Chem Biol 12:694-701 |
| Schmitt, Adam M; Garcia, Julia T; Hung, Tiffany et al. (2016) An inducible long noncoding RNA amplifies DNA damage signaling. Nat Genet 48:1370-1376 |
| Payumo, Alexander Y; Walker, Whitney J; McQuade, Lindsey E et al. (2015) Optochemical dissection of T-box gene-dependent medial floor plate development. ACS Chem Biol 10:1466-75 |
| Bonger, Kimberly M; Rakhit, Rishi; Payumo, Alexander Y et al. (2014) General method for regulating protein stability with light. ACS Chem Biol 9:111-5 |
| Rack, Paul G; Ni, Jun; Payumo, Alexander Y et al. (2014) Arhgap36-dependent activation of Gli transcription factors. Proc Natl Acad Sci U S A 111:11061-6 |
| Cho, Yoon Sun; Jung, Hye Jin; Seok, Seung Hyeok et al. (2013) Functional inhibition of UQCRB suppresses angiogenesis in zebrafish. Biochem Biophys Res Commun 433:396-400 |
| Yamazoe, Sayumi; Shestopalov, Ilya A; Provost, Elayne et al. (2012) Cyclic caged morpholinos: conformationally gated probes of embryonic gene function. Angew Chem Int Ed Engl 51:6908-11 |
| Shestopalov, Ilya A; Pitt, Cameron L W; Chen, James K (2012) Spatiotemporal resolution of the Ntla transcriptome in axial mesoderm development. Nat Chem Biol 8:270-6 |
| Kuo, Alex J; Song, Jikui; Cheung, Peggie et al. (2012) The BAH domain of ORC1 links H4K20me2 to DNA replication licensing and Meier-Gorlin syndrome. Nature 484:115-9 |
