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E-GEOD-78771 - Gene trapping in the Asian malaria vector, Anopheles stephensi

Status
Released on 1 March 2016, last updated on 17 March 2016
Organism
Anopheles stephensi
Samples (1)
Protocols (1)
Description
A piggyBac transposon-based gene trap element was transformed into the Asian malaria vector, Anopheles stephensi, and remobilized using the jumpstarter approach using genetic crosses. Individuals that displayed a gene trap remobilization event were then photodocumented and their RNA and DNA complements were extracted. The DNA compelement was used to determine the genomic insertion site, while the RNA was used to determine the transcript coverage of the genes into which the transposons inserted. In nearly half of the cases, insertion was identified to fall within introns present in the 5'-UTR of transcripts- which are not indicated in the current ab initio models for Anopheles stephensi. The ability to utilize next generation RNA-Seq elucidated the functionality of the gene trap elements that inserted outside of the ab initio gene models, providing clear evidence that not only was the gene trap element working properly, but that it also had a bias towards 5'-end insertion, in particular, 5'-UTR intronic insertion. RNA from 200 pooled individually-extracted An. stephensi that demonstrated a gene trap remobilizable event.
Experiment type
other 
Contacts
William Reid <wzr0005@auburn.edu>, D A O'Brochta, W R Reid
MINSEQE
Exp. designProtocolsVariablesProcessedSeq. reads
Files
Investigation descriptionE-GEOD-78771.idf.txt
Sample and data relationshipE-GEOD-78771.sdrf.txt
Processed data (1)E-GEOD-78771.processed.1.zip
Links