2024/11/25 by Lauren Forbes Beadle, Catherine Sutcliffe, Hilary L. Ashe · 1 voice
Biochemistry, Genetics and Molecular Biology · #Animal Genetics and Reproduction #Biology #CRISPR #CRISPR and Genetic Engineering #Cell biology #Computational biology #Drosophila melanogaster #Gene #Genetics #Genome #RNA and protein synthesis mechanisms #Transcription (linguistics) #Transcription factor #Transposable element
paper · doi:10.1242/dev.204294
openalex created_date 2024/11/25 · openalex publication_date 2024/11/25 · openalex updated_date 2026/08/01
Live imaging of transcription in the Drosophila embryo using the MS2 or PP7 systems is transforming our understanding of transcriptional regulation. However, insertion of MS2/PP7 stem-loops into endogenous genes requires laborious CRISPR genome editing. Here, we exploit the previously described Minos-mediated integration cassette (MiMIC) transposon system in Drosophila to establish a method for simply and rapidly inserting MS2/PP7 cassettes into any of the thousands of genes carrying a MiMIC insertion. In addition to generating a variety of stem-loop donor fly stocks, we have made new stocks expressing the complementary coat proteins fused to different fluorescent proteins. We show the utility of this MiMIC-based approach by MS2/PP7 tagging of endogenous genes and the long non-coding RNA roX1, then imaging their transcription in living embryos. We also present live transcription data from larval brains, the wing disc and ovary, thereby extending the tissues that can be studied using the MS2/PP7 system. Overall, this first high-throughput method for tagging mRNAs in Drosophila will facilitate the study of transcription dynamics of thousands of endogenous genes in a range of Drosophila tissues.