Combinatorial control of gene function with wavelength-selective caged morpholinos.
Caged morpholino oligonucleotides (cMOs) are useful research tools in developmental biology because they allow spatiotemporal control of gene expression in whole organisms. While cMOs are usually triggered by light of a single wavelength, the introduction of spectrally distinct chromophores can enable combinatorial regulation of multiple genes. This chapter describes the general principles and methods of wavelength-selective cMO design and synthesis from commercially available reagents. Synthetic protocols for the linkers and the two-step cMO assembly are described in detail, as well as the microinjection and photoactivation techniques. Following these protocols, spectrally separated cyclic cMOs for multiple genes of interest can be prepared, enabling their inhibition in zebrafish embryos and other animal models.
['Pattanayak|Sankha|S|', 'Vázquez-Maldonado|Luis Angel|LA|', 'Deiters|Alexander|A|', 'Chen|James K|JK|']
D000818:Animals D060326:Chemistry Techniques, Synthetic D020868:Gene Silencing D008027:Light D008845:Microinjections D060172:Morpholinos D016376:Oligonucleotides, Antisense D055668:Photochemical Processes D015027:Zebrafish
Caged morpholinos Developmental biology Zebrafish Antisense oligonucleotides Gene silencing Photolabile linkers Wavelength-selective activation