Detection of Group II Intron-Generated Chimeric mRNAs in Bacterial Cells
Chimeric RNAs are the transcripts composed of exons from two separate genes or transcripts. Although the presence of these joined RNA molecules have mainly been documented in a variety of eukaryotes, we recently demonstrated that the Ll.LtrB group II intron, from the gram-positive bacterium Lactococcus lactis, can generate chimeric mRNAs through a novel intergenic trans-splicing pathway. Here we describe the detailed experimental procedures to detect group II intron-generated mRNA–mRNA chimeras from total RNA extracts using stringent reverse transcription conditions along with a reverse splicing-deficient group II intron as a negative control.
Key wordsGroup II intron Ll.LtrB Bacteria Lactococcus lactis Trans-splicing Chimeric mRNA Circular RNA Circularization
- 8.LaRoche-Johnston F, Monat C, Coulombe S, Cousineau B (2018) Bacterial group II introns generate genetic diversity by circularization and trans-splicing from a population of intron-invaded mRNAs. PLoS Genet 14(11):e1007792. https://doi.org/10.1371/journal.pgen.1007792CrossRefPubMedPubMedCentralGoogle Scholar