ZFIN ID: ZDB-PUB-190803-4
Combinatorial control of gene function with wavelength-selective caged morpholinos
Pattanayak, S., Vázquez-Maldonado, L.A., Deiters, A., Chen, J.K.
Date: 2019
Source: Methods in enzymology   624: 69-88 (Chapter)
Registered Authors: Chen, James K., Pattanayak, Sankha
Keywords: Antisense oligonucleotides, Caged morpholinos, Developmental biology, Gene silencing, Photolabile linkers, Wavelength-selective activation, Zebrafish
MeSH Terms:
  • Animals
  • Chemistry Techniques, Synthetic/methods
  • Gene Silencing*
  • Light
  • Microinjections
  • Morpholinos/administration & dosage
  • Morpholinos/chemical synthesis
  • Morpholinos/chemistry
  • Morpholinos/genetics*
  • Oligonucleotides, Antisense/administration & dosage
  • Oligonucleotides, Antisense/chemical synthesis
  • Oligonucleotides, Antisense/chemistry
  • Oligonucleotides, Antisense/genetics*
  • Photochemical Processes
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed: 31370936 Full text @ Methods Enzymol.
ABSTRACT
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.
ADDITIONAL INFORMATION No data available