An RNA biosensor for imaging the first round of translation from single cells to living animals

Halstead, James M.; Lionnet, Timothée; Wilbertz, Johannes; Wippich, Frank; Ephrussi, Anne; Singer, Robert H.; Chao, Jeffrey A.

doi:10.1126/science.aaa3380

Cited by 252 publications

(246 citation statements)

References 37 publications

Supporting

Mentioning

237

Contrasting

Order By: Relevance

“…We assessed this possibility by visualizing translation within SICs after treatment with puromycin -an antibiotic that is incorporated into elongating polypeptides, resulting in the premature release of truncated puromycinylated polypeptides (Schmidt et al, 2009). Although alternative methods could have been used for localized translation visualization (Fallini et al, 2016;Griffin et al, 1998;Halstead et al, 2015), only puromycin incorporation offers a rapid and precise glimpse of translation events, making it a well-suited method to assess translation events in rapid biological processes, such as cell adhesion. Hence, MRC-5 cells were allowed to adhere for 60 min, prior to the addition of 5 μg ml −1 puromycin to the medium 5 min before fixation.…”

Section: Localized Mrna Translation Within Sicsmentioning

confidence: 99%

Localized translation regulates cell adhesion and transendothelial migration

Bergeman¹,

Caillier²,

Houle³

et al. 2016

Development

View full text Add to dashboard Cite

Epithelial-to-mesenchymal transition (EMT) is a process by which cancer cells gain the ability to leave the primary tumor site and invade surrounding tissues. These metastatic cancer cells can further increase their plasticity by adopting an amoeboid-like morphology, by undergoing mesenchymal-to-amoeboid transition (MAT). We found that adhering cells produce spreading initiation centers (SICs), transient structures that are localized above nascent adhesion complexes, and share common biological and morphological characteristics associated with amoeboid cells. Meanwhile, spreading cells seem to return to a mesenchymal-like morphology. Thus, our results indicate that SIC-induced adhesion recapitulates events that are associated with amoeboid-to-mesenchymal transition (AMT). We found that polyadenylated RNAs are enriched within SICs, blocking their translation decreased adhesion potential of metastatic cells that progressed through EMT. These results point to a so-farunknown checkpoint that regulates cell adhesion and allows metastatic cells to alter adhesion strength to modulate their dissemination.

show abstract

Section: Localized Mrna Translation Within Sicsmentioning

confidence: 99%

Localized translation regulates cell adhesion and transendothelial migration

Bergeman¹,

Caillier²,

Houle³

et al. 2016

Development

View full text Add to dashboard Cite

show abstract

“…The most common technique involves integrating multiple copies of an RNA stem-loop sequence into the target transcript that binds MS2 bacteriophage coat protein with high-affinity. [136][137][138] The MS2 has proven powerful for large transcripts but smaller RNAs such as siRNAs and miRNAs cannot be visualized with this technique because it would require too large a tag. Minimal systems that might fit the loop region of a hairpin would be a valuable innovation.…”

Section: Chemical Approaches Toward Sequence Selective Nucleic Acid Amentioning

confidence: 99%

Properties and reactivity of nucleic acids relevant to epigenomics, transcriptomics, and therapeutics

2016

View full text Add to dashboard Cite

Developments in epigenomics, toxicology, and therapeutic nucleic acids all rely on a precise understanding of nucleic acid properties and chemical reactivity. In this review we discuss the properties and chemical reactivity of each nucleobase and attempt to provide some general principles for nucleic acid targeting or engineering. For adenine-thymine and guaninecytosine base pairs, we review recent quantum chemical estimates of their Watson-Crick interaction energy, - stacking energies, as well as the nuclear quantum effects on tautomerism. Reactions that target nucleobases have been crucial in the development of new sequencing technologies and we believe further developments in nucleic acid chemistry will be required to deconstruct the enormously complex transcriptome.

show abstract

“…Resolving RNA and protein localizations is particularly important for complex neuronal structures in which dendritic and axonal processes may span hundreds to thousands of cell bodies away from the soma. Here, local rates of translation and RNA/protein modifications are determined by local signalling mechanisms rather than necessarily the transcriptional state of the cell [53,56,57]. In situ characterization of cell and synapse sub-types should again prove highly useful to informing structure-based models of neuronal circuit function.…”

Section: Multiplexed Imaging To Resolve Distinct Cellular and Moleculmentioning

confidence: 99%

Advancing multiscale structural mapping of the brain through fluorescence imaging and analysis across length scales

Hogstrom¹,

Guo²,

Murugadoss³

et al. 2016

Interface Focus.

View full text Add to dashboard Cite

Brain function emerges from hierarchical neuronal structure that spans orders of magnitude in length scale, from the nanometre-scale organization of synaptic proteins to the macroscopic wiring of neuronal circuits. Because the synaptic electrochemical signal transmission that drives brain function ultimately relies on the organization of neuronal circuits, understanding brain function requires an understanding of the principles that determine hierarchical neuronal structure in living or intact organisms. Recent advances in fluorescence imaging now enable quantitative characterization of neuronal structure across length scales, ranging from single-molecule localization using super-resolution imaging to whole-brain imaging using light-sheet microscopy on cleared samples. These tools, together with correlative electron microscopy and magnetic resonance imaging at the nanoscopic and macroscopic scales, respectively, now facilitate our ability to probe brain structure across its full range of length scales with cellular and molecular specificity. As these imaging datasets become increasingly accessible to researchers, novel statistical and computational frameworks will play an increasing role in efforts to relate hierarchical brain structure to its function. In this perspective, we discuss several prominent experimental advances that are ushering in a new era of quantitative fluorescence-based imaging in neuroscience along with novel computational and statistical strategies that are helping to distil our understanding of complex brain structure.

show abstract

An RNA biosensor for imaging the first round of translation from single cells to living animals

Cited by 252 publications

References 37 publications

Localized translation regulates cell adhesion and transendothelial migration

Localized translation regulates cell adhesion and transendothelial migration

Properties and reactivity of nucleic acids relevant to epigenomics, transcriptomics, and therapeutics

Advancing multiscale structural mapping of the brain through fluorescence imaging and analysis across length scales

Contact Info

Product

Resources

About