Antisense oligonucleotides (ASO) represent a promising approach for future clinical applications. These 16-20nt single-stranded DNA or RNA molecules show the capability to pass the plasma membrane, enter the cellular nucleus and pair with complementary mRNA. This recruits duplex-RNA digesting enzymes (e.g., RNAseH1), which eliminate the respective RNA. Another mechanism of ASO action consists in the modulation of RNA splicing.
Little is known about the pathway by which ASOs contact the cellular surface, enter the cell, escape from importing vehicles (like endocytic vesicles) and enter the nucleus.
In this work I am generating a reporter cell line to measure ASO efficiency. The advent of CRISPR/Cas9 mediated gene silencing and genome-scale sgRNA libraries allows us to screen reporter-expressing cells for molecular factors governing the activity of antisense oligonucleotides.
Institute of pharmacology, Medical University of Vienna, Austria.