Previously, on the story of MS2 in yeast: Last year, Roy Parker published a short article, in which he claimed that using the MS2 system in yeast causes the accumulation of 3′ RNA fragments, probably due to inhibition of mRNA degradation by the 5′ to 3′ exoribonuclease Xrn1. He argued that these findings put in question all the work on mRNA localization in yeast using the MS2 system. About a year later, we wrote a response to that article. We argued that, yes, such fragments exist, but 1. most of it stems from over-expression of the labeled mRNA. Parker agreed with that. 2. That these fragments accumulate in P-bodies, and are distinguishable from single mRNAs and we can discard cells which show these structures. 3. We argued that this might not be the case for every mRNA and should be tested on a case by case basis. 4. We and Parker agreed that the best way to determine if such fragments exist is by performing single-molecule FISH (smFISH) with double labeling – a set of probes for the length of the mRNA and a set of probes for the MS2 stem-loops. Now, a new paper from Karsten Weis’ lab shows more evidence, by doing smFISH, for the existence of these fragments.
Most recent posts:
Posts are categorized as follows:
Tags3D-SIM ASAPbio ascb15 Bacteria BFP Bio-protocol BioimageXD bioRxiv blinking Brain brightness CASFISH CellProfiler CFP Channelrhodopsin Chicken Chrimson Chronos circularly permutated CLARITY clover CRISPR CSHL cy3 CyTOF DAPI definitions DiD DiL DiO DNA FISH dTomato E. coli EC EL222 emerald EoS ER ER-tracker ERK exosomes EYFP Fiji FISEB Fish (the animal) FISH-Quant FISSEQ FITC FlAsH flu Fly FM dyes folding FUNCAT gaussian fit GCaMP GFP GFPm GFPmut2 GFPmut3 glutamate sensor GRC HaloTag HHMI Janelia histones HIV(AIDS) Hoecht Icy ImageJ imaging_at_home ISCR Its broken! IVF Kinesin LanYFP laser LSSmKate LSSmOrange Lysotracker Malaria Mammalian cell maturation mCherry mCLING membrane nanotubes microvesicles mitochondria MitoTracker mKate2 mNeonGreen mRNA decay mRNA export mRNA localization MS2 mutations my pics nanobodies Nat Methods neuraminidase neurons NIH Image Nobel prize not imaging NPC oligomerization omnidirectional oocyte PALM PAmCherry peroxisomes personal experience pHluorin photoinhibition photostability phototoxicity pHTomato Pol II CTD PP7 probe design propidiume iodide QCBNet Qdot625 Qdot655 quantitative microscopy QY RCas9 ReaChr receptor internalization resolution RFP Rhodamine 123 RNAScope Rodent science career shutter SiNC Singer lab single cell RNA-seq single molecule SmartFlare spaghetti monster spectrum SPIM Spinach split-GFP spot analysis STORM stress granules Suntag super-resolution supercharger superfast superfolder superglo TagRFP657 tenure track tethering tetramethylrosamine tFT TGF-beta transcription translation TRICK tRNA UnaG venus Worm yeast zebra fish zygote
Archive of Green Fluorescent Blog
Help me attend a conference