A flexible method to image unmodified transcripts and transcription in vivo would be a valuable tool to understand the regulation and dynamics of transcription. Here, we present a novel approach to follow native transcription, with fluorescence microscopy, in live C. elegans. By using the fluorescently tagged Argonaute protein NRDE-3, programmed by exposure to defined dsRNA to bind to nascent transcripts of the gene of interest, we demonstrate transcript labelling of multiple genes, at the transcription site and in the cytoplasm. This flexible approach does not require genetic manipulation, and can be easily scaled up by relying on whole-genome dsRNA libraries. We apply this method to image the transcriptional dynamics of the heat-shock inducible gene hsp-4 (a member of the hsp70 family), as well as two transcription factors: ttx-3 (a LHX2/9 orthologue) in embryos, and hlh-1 (a MyoD orthologue) in larvae, respectively involved in neuronal and muscle development.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8421136 | PMC |
| http://dx.doi.org/10.1093/nar/gkab469 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Diabetes Management Using a Patient Navigator in a Native Hawaiian Population: Experiences and Perceptions of the Kilolani Project.
Int J Environ Res Public Health
January 2025
The Queen's Health System, Honolulu, HI 96813, USA.
Native Hawaiians (NHs) are a historically oppressed population disproportionately burdened by diabetes and related complications. The Kilolani Project, a patient navigator-centered, chronic disease management program, targets upstream drivers of health among vulnerable NH adult patients with diabetes within an urban academic safety-net clinic. To investigate the impact of the Kilolani Project, we performed a qualitative study to examine patient perspectives.
View Article and Find Full Text PDFHighly multiplexed spatial transcriptomics in bacteria.
Science
January 2025
Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
Single-cell decisions made in complex environments underlie many bacterial phenomena. Image-based transcriptomics approaches offer an avenue to study such behaviors, yet these approaches have been hindered by the massive density of bacterial messenger RNA. To overcome this challenge, we combined 1000-fold volumetric expansion with multiplexed error-robust fluorescence in situ hybridization (MERFISH) to create bacterial-MERFISH.
View Article and Find Full Text PDFIdentification of the bHLH gene family and functional analysis of ChMYC2 in drought stress of Cerasus humilis.
Plant Physiol Biochem
January 2025
Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast Forestry University), Ministry of Education, Harbin, 150040, China; College of Life Science, Northeast Forestry University, Harbin, 150040, China. Electronic address:
The basic helix-loop-helix (bHLH) transcription factors (TFs) play a crucial regulatory role in the growth and development of plants, as well as in their response to environmental stresses. In this study, we identified 94 ChbHLHs from Cerasus humilis, an economically valuable tree native to northern China. We analyzed their evolutionary relationships, gene structures, chromosome distributions, promoter cis-regulatory elements, and collinearity.
View Article and Find Full Text PDFA practical guide for choosing an optimal spatial transcriptomics technology from seven major commercially available options.
BMC Genomics
January 2025
UCLA Technology Center for Genomics & Bioinformatics, Department of Pathology & Laboratory Medicine, 650 Charles E Young Dr. South, Los Angeles, CA, 90095, USA.
Spatial transcriptomics technology enables the mapping of gene expression within tissues, allowing researchers to visualize the spatial distribution of RNA molecules and gain insights into cellular organization, interactions, and functions in their native environments. A variety of spatial technologies are now commercially available, each offering distinct technical parameters such as cellular resolution, detection sensitivity, gene coverage, and throughput. This wide range of options can make it challenges or create confusion for researchers to select the most appropriate platform for their specific research objectives.
View Article and Find Full Text PDFReal-time visualization of reconstituted transcription reveals RNA polymerase II activation mechanisms at single promoters.
bioRxiv
January 2025
Dept. of Biochemistry, University of Colorado, Boulder, CO, 80303, USA.
RNA polymerase II (RNAPII) is regulated by sequence-specific transcription factors (TFs) and the pre-initiation complex (PIC): TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH, Mediator. TFs and Mediator contain intrinsically-disordered regions (IDRs) and form phase-separated condensates, but how IDRs control RNAPII function remains poorly understood. Using purified PIC factors, we developed a Real-time In-vitro Fluorescence Transcription assay (RIFT) for second-by-second visualization of RNAPII transcription at hundreds of promoters simultaneously.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!