Technology breakthrough reveals cellular transcription process

December 4, 2014, University of Montreal

A new technology that reveals cellular gene transcription in greater detail has been developed by Dr. Daniel Kaufmann of the University of Montreal Hospital Research Centre (CRCHUM) and the research team he directed. "This new research tool offers us a more profound view of the immune responses that are involved in a range of diseases, such as HIV infection. At the level of gene transcription, this had been difficult, complex and costly to do with current technologies, such as microscopy," explained the University of Montreal professor.

The technology is known as the "FISH investigation protocol" (Fluorescent in situ hybridization) - it optimises current flow cytometry processes, resulting in a greater visibility of the transcription of RNA, a key step in the transmission of cellular information. The researchers published their breakthrough in Nature Communications' online edition. Flow cytometry itself enables the simultaneous measurement of hundreds of thousands of cells as they quickly pass through laser beams. This technology dates back to 1934, when it was used to count the cells making up organs and blood, before being developed across the decades to enable the analysis of cells' various physical characteristics. Today, flow cyclometers are standard equipment for immunology and cancer research labs.

However, the tool's major barrier was that it could only be used for the study of proteins, as specific antibodies that recognize these molecules make them visible by marking. Unfortunately, there are many proteins for which no specific antibodies are available, and so it has been impossible to study them with flow cytometry. Moreover, cancer, HIV and other diseases are caused by a problem that precedes cells' creation of proteins - the deregulation of the of DNA information to the RNA messenger. Classic cannot examine what is happening during this process.

Dr Kaufmann's team worked with Affymetrix, a company, to break through this barrier by applying their technology to a fundamental step in the transmission of genetic information within the cell. The technology, which is now patented, involves carefully mixing chemical reactants by following a two day long handling protocol. Using a standard flow cyclometer, a hybridization oven and a variety of meticulously dosed solutions (which act as microscopic fluorescent probes), the cells can be coaxed to deliver information that has been previously unavailable.

The application of the technology promises to have a huge impact as ultimately it will enable researchers to better understand the mechanisms that cause diseases and to establish the efficacy of the drugs used to treat them. "In principle, our technology can be applied to any species. We can detect a very wide variety of RNA molecules produced by the human body and animals," Dr. Kaufmann explained.

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JVK
1 / 5 (2) Dec 04, 2014
High-throughput detection of miRNAs and gene-specific mRNA at the single-cell level by flow cytometry http://www.nature...641.html

Excerpt: "We use this technique to show modulation of a microRNA critical for T-cell function, ​miR-155. We adapt this assay for simultaneous detection of mRNA and proteins by ImageStream technology."

I've requested a reprint and hope they linked nutrient-dependent RNA-directed DNA methylation from RNA-mediated events and amino acid substitutions to cell type differentiation via epigenetic effects on the microRNA/messenger RNA balance in species from microbes to man.

For examples see: Nutrient-dependent/pheromone-controlled adaptive evolution: a model http://www.ncbi.n...24693353

Excerpt: "..the epigenetic 'tweaking' of the immense gene networks that occurs via exposure to nutrient chemicals and pheromones can now be modeled in the context of the microRNA/messenger RNA balance..."
JVK
1 / 5 (2) Dec 04, 2014
http://phys.org/n...html#jCp

"The researchers then investigated how the history and ecology affected speciation among the 27 lineages of birds. They discovered the longer length of time a species can inhabit an area, the more likely it will disperse and diverge. Also, the less mobility a species has, the more likely it will diverge as well."

Ongoing discussion among science idiots (Ren82 excepted) has continued to ignore the obvious role that nutrient-dependent changes in the microRNA/messenger RNA balance play in cell type differentiation via amino acid substitutions.

If you are not an evolutionary theorist and have not been taught to believe in the pseudoscientific nonsense of their ridiculous theories, you might enjoy the comments made by anonymous fools and idiot minions of biology teachers like PZ Myers -- if only for comparison to what you understand about biologically-based facts.
Captain Stumpy
5 / 5 (2) Dec 04, 2014
Ongoing discussion among science idiots
you are still sore because you have been proven wrong by Anonymous and myself time and again, especially with regard to your creationist religious belief that all mutations are harmful

you ignore evidence that you don't like, especially since the author called you out and said you not only LIED about the work, but didn't understand it: http://myxo.css.m...dex.html
http://www.extavourlab.com/
PLUS you still cant understand that your own model causes mutations, even though you already admitted it!
remember when i asked
DOES your model make any changes to the nucleotide sequence of the genome of an organism, virus, or extrachromosomal genetic element?
This is a yes or no answer
(this is the DEFINITION of mutation) to which you answered
YES!
--Thanks for asking
your own words, not mine
you are proven to be a liar, fool and unable to comprehend modern biology

AND worse yet, you are a creationist TROLL
JVK
1 / 5 (2) Dec 04, 2014
proven wrong by Anonymous and myself [Captain Stumpy]


Their assay simultaneously detects changes in the epigenetically-effected miRNA/mRNA balance and proteins, which link nutrient-uptake to the physical landscape of DNA in the organized genomes of species from microbes to man.

At the single-cell level, they are beginning to assess how RNA-directed DNA methylation and RNA-mediated events link bio-physically constrained thermodynamic cycles of protein biosynthesis and degradation to "...translational and expressional effects in both normal and pathologic conditions."

They have moved beyond what already is available to link metabolic networks and genetic networks in the context of pharmacogenomics, which enabled serious scientists to distinguish the difference between effects of mutations that perturb protein folding and amino acid substitutions that stabilize DNA in organized genomes via fixation in the context of pheromone-controlled reproduction.
JVK
1 / 5 (2) Dec 04, 2014
I'm not sure what aspect of their assay others think proves I am wrong about anything portrayed in my series of published works, which began in 1996 with "From Fertilization to Adult Sexual Behavior"

http://www.hawaii...ion.html

In our section on Molecular epigenetics, we (TB) wrote:

"Small intranuclear proteins also participate in generating alternative splicing techniques of pre-mRNA and, by this mechanism, contribute to sexual differentiation in at least two species..."

What we now see is a report on an assay that shows alternative splicing techniques of pre-mRNA may lead to cell type differentiation of all cell types in all individuals of all species via conserved molecular mechanisms of epigenetically-effected self vs other recognition in the context of feedback loops that link nutrient-uptake to the pheromone-controlled physiology of reproduction.

There's a model for that! http://youtu.be/DbH_Rj9U524
Captain Stumpy
5 / 5 (2) Dec 05, 2014
I'm not sure what aspect of their assay others think proves I am wrong about anything portrayed in my series of published works
logical fallacy- appeal to authority

except, of course, you are not one, and you blatantly lie

you cannot even properly utilize the nomenclature (lexicon) of the field of Biology/Genetics as you completely ignore the definition of mutation

and then your constant denigration of beneficial mutation causes most to believe you are being blatantly stupid as you have been corrected repeatedly, especially as you then turn around and promote it with your model, as shown above and here: http://phys.org/n...rge.html

you ignore empirical evidence:
http://myxo.css.m...dex.html
http://www.extavourlab.com/

you promote creationist religion while it has been demonstrated that creationists have no science in their movement:
https://en.wikipe...Arkansas
OZGuy
5 / 5 (2) Dec 05, 2014
Closing paragraph from: http://www.scient...es-real/

"The failure to identify human pheromones has not stopped some enterprising individuals from trying to make a profit from love potions purporting to contain pheromones. In reality, these products often use pig pheromones. "They don't have any history in the biomedical literature—they just fell out of the sky," says olfactory neuroscientist Charles Wysocki, also of Monell. For now, the idea of perfumes and potions based on human pheromonal communication just doesn't pass the sniff test."
JVK
1 / 5 (2) Dec 05, 2014
Excerpted from this article:
Dr Kaufmann's team worked with Affymetrix, a company, to break through this barrier by applying their technology to a fundamental step in the transmission of genetic information within the cell.


They claim: "High-throughput detection of miRNAs and gene-specific mRNA at the single-cell level by flow cytometry"

I claim: "..the epigenetic 'tweaking' of the immense gene networks that occurs via exposure to nutrient chemicals and pheromones can now be modeled in the context of the microRNA/messenger RNA balance..."

An anonymous fool infers that Wysocki's claim is pertinent to discussion of the obvious link from the epigenetic landscape to the physical landscape of DNA in the organized genomes of species from microbes to man.

Closing sentence from this post: Anonymous fools invariably are science idiots!
JVK
1 / 5 (2) Dec 05, 2014
Re:
They don't have any history in the biomedical literature...


Effects of 5alpha-androst-16-en-3alpha-ol on the pulsatile secretion of luteinizing hormone in human females http://chemse.oxf...465.long

"the present results suggest that [5alpha-androst-16-en-3alpha-ol] 3alpha-androstenol may be a pheromone included in the axillary compounds..."

Axillary pheromones modulate pulsatile LH secretion in humans http://www.ncbi.n...11303754 "The frequency of the LH pulse was increased by FP compounds and was decreased by OP compounds..."

Feedback loops link odor and pheromone signaling with reproduction http://www.scienc...05009815

"Indications that GnRH peptide plays an important role in the control of sexual behaviors suggest that pheromone effects on these behaviors might also involve GnRH neurons."

GnRH pulse frequency modulates LH.

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