Plenary Session 4 – Chemical Tools
9/1 11:00 - 1:00, Memorial Auditorium
A caged morpholino-based strategy for transcriptional target discovery
James K. Chen
Deciphering the molecular mechanisms that regulate vertebrate development and physiology requires an ability to alter these genetic programs with spatiotemporal precision. Caged morpholino (cMO) oligonucleotides enable light-controlled gene silencing in zebrafish and other optically transparent organisms, and we report here the application of cMOs to discover transcriptional targets of the T-box gene no tail (ntl). In contrast to previous whole-embryo analyses of the ntl-dependent transcriptome, we have identified ntl targets in a tissue-specific manner by combining cMOs, photoactivatable fluorophores, fluorescence-activated cell sorting, and microarrays. Using this strategy, we have discovered distinct sets of ntl-regulated genes that contribute differentially to notochord formation and maturation.
University of North Carolina.
University of California, San Francisco.