A global genetic analysis of nematode spliced-leader trans-splicing
Jonathan PettittBerndt MuellerBernadette Connolly
Spliced-leader trans-splicing
‘Slintron’
Taxonomic Distribution of SL trans-splicing
Origin ofSpliced-Leader trans-splicing
C. elegansSL1 RNA
U1 snRNA
TMG
TMG
SM
SM
SL trans-splicing in C. elegans• 70% of C. elegans genes are
SL1 trans-spliced• Loss of SL1 RNA gene is
embryonic lethal• Many questions remain
– how are trans-splicing and cis-splicing coordinated in the same cell?
– How is the SL RNA brought to the 5’end of the mRNA?
– How are SM proteins recycled from the SL introns?
An in vivo assay for SL trans-splicing
GFPSL1
GFPATG
outron
SL trans-splicing
Screening for molecules required for SL trans-splicing
GFPATG
outron
Control mRNA
Candidate RNAi Screens
Summary
• Developed in vivo assay to visualise defects in SL trans-splicing
• First ever systematic genetic screens for molecules required for SL trans-splicing in any organism
• SL trans-splicing an ideal target for the development of novel anti-helminthics:– Not present in plants or vertebrates