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A global genetic analysis of nematode spliced-leader trans-splicing
Jonathan PettittBerndt MuellerBernadette Connolly
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Spliced-leader trans-splicing
‘Slintron’
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Taxonomic Distribution of SL trans-splicing
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Origin ofSpliced-Leader trans-splicing
C. elegansSL1 RNA
U1 snRNA
TMG
TMG
SM
SM
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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?
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An in vivo assay for SL trans-splicing
GFPSL1
GFPATG
outron
SL trans-splicing
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Screening for molecules required for SL trans-splicing
GFPATG
outron
Control mRNA
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Candidate RNAi Screens
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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