affect nearly a billion people living in low income countries around the world.
drugs has resulted in emergence of anthelminthic resistant parasitic nematodes.
parasites that are resistant to single drugs, and even to multi-drug combinations.
As such, there is an urgent need to
develop new anthelmintic drugs with novel modes of action. One potential source
of targets is spliced leader (SL1)
trans-splicing, an essential gene expression
step that is unique to nematodes. We
have adapted an
in vivo green fluorescent protein (GFP) based assay of
SL1
trans-splicing in
Caenorhabditis elegans which is suitable
for ultrahigh-throughput screening (uHTS) in the search for novel anthelminthic
drugs. Using sinefungin, a known inhibitor of SL
trans-splicing, as a positive chemical control, a robust HTS
protocol was established combining automated dispensing of
C. elegansworms into 384- or 1536- well plates with addition of compounds using an
acoustic liquid dispenser and the detection of the inhibition of SL
trans-splicing
in a standard multi-label microplate reader. A pilot screen was performed to
validate the utility of the protocol to screen compounds.
References:
1. Pandarakalam,
G., Speake, M., McElroy, S., Alturkistani, A., Philippe, L., Pettitt, J.,
Müller, B. and Connolly, B. (2019). A high-throughput screen for the
identification of compounds that inhibit nematode gene expression by targeting
spliced leader trans-splicing. International Journal for
Parasitology: Drugs and Drug Resistance, 10, pp.28-37.
2. Philippe,
L., Pandarakalam, G., Fasimoye, R., Harrison, N., Connolly, B., Pettitt, J. and
Müller, B. (2017). An in vivo genetic screen for genes involved in spliced
leader trans-splicing indicates a crucial role for continuous de novo
spliced leader RNP assembly. Nucleic Acids Research, 45(14),
pp.8474-8483.
