Conditional control of alternative splicing through light-triggered splice-switching oligonucleotides.
The spliceosome machinery is composed of several proteins and multiple small RNA molecules that are involved in gene regulation through the removal of introns from pre-mRNAs in order to assemble exon-based mRNA containing protein-coding sequences. Splice-switching oligonucleotides (SSOs) are genetic control elements that can be used to specifically control the expression of genes through correction of aberrant splicing pathways. A current limitation with SSO methodologies is the inability to achieve conditional control of their function paired with high spatial and temporal resolution. We addressed this limitation through site-specific installation of light-removable nucleobase-caging groups as well as photocleavable backbone linkers into synthetic SSOs. This enables optochemical OFF → ON and ON → OFF switching of their activity and thus precise control of alternative splicing. The use of light as a regulatory element allows for tight spatial and temporal control of splice switching in mammalian cells and animals.
†Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.|‡Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
James
J
Hemphill
‡Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
Qingyang
Q
Liu
‡Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
Rajendra
R
Uprety
†Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.
Subhas
S
Samanta
§Department of Developmental Biology, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania 15260, United States.
Michael
M
Tsang
∥Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, United States.
Rudolph L
RL
Juliano
†Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.|‡Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
Alexander
A
Deiters
D017398:Alternative Splicing / Q000528:radiation effects*; D000818:Animals; D006367:HeLa Cells; D006801:Humans; D008027:Light; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics*; D015027:Zebrafish
D009841:Oligonucleotides