Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

New Tool Isolates RNA within Specific Cells

19.05.2009
A team of University of Oregon biologists, using fruit flies, has created a way to isolate RNA from specific cells, opening a new window on how gene expression drives normal development and disease-causing breakdowns.

While DNA (deoxyribonucleic acid) provides an identical genetic blueprint in every cell, RNA (ribonucleic acid) decodes genetic instructions that turn protein molecules on and off in different cell types.

The new tagging method, tested in a variety of subsets of Drosophila brain cells, is described in a paper put on line ahead of regular publication by the journal Nature Methods. Instead of scientists needing to physically separate cell types, they now can inject a chemically modified gene from the one-celled organism Toxoplasma gondii and activate it in only one cell type within a tissue. Only newly generated RNA in this cell type is then tagged and isolated.

"By analyzing RNA from different cell types, we can begin to understand how cellular differences are generated," said lead author Michael R. Miller, a National Science Foundation-funded doctoral student in the lab of Chris Doe, a UO biologist and Howard Hughes Medical Institute (HHMI) investigator. "Our new TU-tagging method should be useful for isolating cell-type specific RNA from other organisms, including mammals, and should be useful in broad areas of research including studies of development, neurobiology and disease."

The new non-toxic, non-invasive method makes it possible to "listen in" to the messages -- in fact, messenger RNA -- that the nucleus is sending each cell, without perturbing the cell, Doe said. "It is much like eavesdropping on a phone conversation, rather than pulling the person out of the house for questioning. The cell has no idea that its RNAs are being 'tagged' for isolation and study. That's good, because we get a more accurate idea of what the cell is saying."

That, Doe added, could be helpful for 'listening' to host cells before and after the initiation of a disease to determine how cells respond, or, for example study healthy immune cells versus bacterially-challenged immune cells or neurons before they learn a task and after they learn a task to determine what changes in the cell are caused by the experience.

The new UO-developed tool builds on work led by co-author Michael D. Cleary, who as a doctoral student at Stanford University unveiled the T. gondii-based approach for use in analyzing RNA synthesis and decay in 2005 in Nature Biotechnology. Cleary, now a faculty member at the University of California, Merced, worked on the UO project as a postdoctoral fellowship funded by the National Institutes of Health and HHMI.

Cleary's group built its tool with the enzyme uracil phosphoribosyltransferase (UPRT), a nucleotide salvage enzyme that prepares nucleotides for incorporation into newly synthesized RNA. By altering the nucleotide analog 4-thiouracil, the UPRT enzyme caused RNA to become tagged with thiouracil (TU), allowing the "TU-tagged" RNA to be purified from untagged RNA.

In Doe's lab, Miller, Cleary and research technician Kristin J. Robinson of the UO's institutes of Neuroscience and Molecular Biology manipulated Drosophila so that they would only express UPRT in specific target cells. The group tested the new approach in embryos, larvae and adults using microarray technology to detect cell type-specific gene expression. The researchers say the method should work in other systems, including vertebrates, by using gene transfer, retroviral delivery, electrical pulses of molecules through cell membranes, or messenger RNA injection.

About the University of Oregon

The University of Oregon is a world-class teaching and research institution and Oregon's flagship public university. The UO is a member of the Association of American Universities (AAU), an organization made up of the 62 leading public and private research institutions in the United States and Canada. The University of Oregon is one of only two AAU members in the Pacific Northwest.

Video: http://www.youtube.com/watch?v=IFTYrI048uE

Sources: Chris Doe, professor of biology and HHMI investigator, 541-346-4877, cdoe@uoregon.edu; Michael R. Miller, UO doctoral student, mmiller@uoregon.edu

Jim Barlow | Newswise Science News
Further information:
http://www.uoregon.edu

More articles from Life Sciences:

nachricht Rainbow colors reveal cell history: Uncovering β-cell heterogeneity
22.09.2017 | DFG-Forschungszentrum für Regenerative Therapien TU Dresden

nachricht The pyrenoid is a carbon-fixing liquid droplet
22.09.2017 | Max-Planck-Institut für Biochemie

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: The pyrenoid is a carbon-fixing liquid droplet

Plants and algae use the enzyme Rubisco to fix carbon dioxide, removing it from the atmosphere and converting it into biomass. Algae have figured out a way to increase the efficiency of carbon fixation. They gather most of their Rubisco into a ball-shaped microcompartment called the pyrenoid, which they flood with a high local concentration of carbon dioxide. A team of scientists at Princeton University, the Carnegie Institution for Science, Stanford University and the Max Plank Institute of Biochemistry have unravelled the mysteries of how the pyrenoid is assembled. These insights can help to engineer crops that remove more carbon dioxide from the atmosphere while producing more food.

A warming planet

Im Focus: Highly precise wiring in the Cerebral Cortex

Our brains house extremely complex neuronal circuits, whose detailed structures are still largely unknown. This is especially true for the so-called cerebral cortex of mammals, where among other things vision, thoughts or spatial orientation are being computed. Here the rules by which nerve cells are connected to each other are only partly understood. A team of scientists around Moritz Helmstaedter at the Frankfiurt Max Planck Institute for Brain Research and Helene Schmidt (Humboldt University in Berlin) have now discovered a surprisingly precise nerve cell connectivity pattern in the part of the cerebral cortex that is responsible for orienting the individual animal or human in space.

The researchers report online in Nature (Schmidt et al., 2017. Axonal synapse sorting in medial entorhinal cortex, DOI: 10.1038/nature24005) that synapses in...

Im Focus: Tiny lasers from a gallery of whispers

New technique promises tunable laser devices

Whispering gallery mode (WGM) resonators are used to make tiny micro-lasers, sensors, switches, routers and other devices. These tiny structures rely on a...

Im Focus: Ultrafast snapshots of relaxing electrons in solids

Using ultrafast flashes of laser and x-ray radiation, scientists at the Max Planck Institute of Quantum Optics (Garching, Germany) took snapshots of the briefest electron motion inside a solid material to date. The electron motion lasted only 750 billionths of the billionth of a second before it fainted, setting a new record of human capability to capture ultrafast processes inside solids!

When x-rays shine onto solid materials or large molecules, an electron is pushed away from its original place near the nucleus of the atom, leaving a hole...

Im Focus: Quantum Sensors Decipher Magnetic Ordering in a New Semiconducting Material

For the first time, physicists have successfully imaged spiral magnetic ordering in a multiferroic material. These materials are considered highly promising candidates for future data storage media. The researchers were able to prove their findings using unique quantum sensors that were developed at Basel University and that can analyze electromagnetic fields on the nanometer scale. The results – obtained by scientists from the University of Basel’s Department of Physics, the Swiss Nanoscience Institute, the University of Montpellier and several laboratories from University Paris-Saclay – were recently published in the journal Nature.

Multiferroics are materials that simultaneously react to electric and magnetic fields. These two properties are rarely found together, and their combined...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

“Lasers in Composites Symposium” in Aachen – from Science to Application

19.09.2017 | Event News

I-ESA 2018 – Call for Papers

12.09.2017 | Event News

EMBO at Basel Life, a new conference on current and emerging life science research

06.09.2017 | Event News

 
Latest News

Penn first in world to treat patient with new radiation technology

22.09.2017 | Medical Engineering

Calculating quietness

22.09.2017 | Physics and Astronomy

Hope to discover sure signs of life on Mars? New research says look for the element vanadium

22.09.2017 | Physics and Astronomy

VideoLinks
B2B-VideoLinks
More VideoLinks >>>