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