Citation:
Curr Protoc Nucleic Acid Chem. 2015;62:4.65.1-4.65.26. doi: 10.1002/0471142700.nc0465s62
Abstract:
Phosphorodiamidate morpholino oligomers (PMOs) are a highly capable class of synthetic antisense oligonucleotides that are used to study gene functions in in vitro and in vivo models. This unit describes the synthesis of exocyclic-amine-protected 7′-hydroxy and 7′-chlorophosphoramidate-activated morpholino monomers of A, T, G, and C, together with their incorporation into short PMO oligomers by solid-phase synthesis. Starting from ribonucleosides, the exocyclic-amine-protected 7′-hydroxy monomers are prepared following a modified Summerton protocol, which consists of a periodate cleavage/Schiff base formation/reduction cycle. The exocyclic amine protections are installed at a later stage (except G) to avoid the use of costly exocyclic-amine-protected counterparts that give control over protecting group manipulation. The 7′-hydroxy monomers with N-Trit/N-MMTr are then converted to the 7′-chlorophosphoramidate morpholino monomers in one step employing a combination of lithium bromide and DBU. These chlorophosphoramidate monomers are finally assembled by solid-support synthesis to obtain the short PMO oligomers.
Epub:
Not Epub
Link to Publication:
http://onlinelibrary.wiley.com/doi/10.1002/0471142700.nc0465s62/abstract