Navigation Links
Researchers develop new way to see single RNA molecules inside living cells
Date:4/6/2009

Biomedical engineers have developed a new type of probe that allows them to visualize single ribonucleic acid (RNA) molecules within live cells more easily than existing methods. The tool will help scientists learn more about how RNA operates within living cells.

Techniques scientists currently use to image these transporters of genetic information within cells have several drawbacks, including the need for synthetic RNA or a large number of fluorescent molecules. The fluorescent probes developed at the Georgia Institute of Technology circumvent these issues.

"The probes we designed shine bright, are small and easy to assemble, bind rapidly to their targets, and can be imaged for hours. These characteristics make them a great choice for studying the movement and location of RNA inside a single cell and the interaction between RNA and binding proteins," said Philip Santangelo, an assistant professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University.

Details of the probe production process and RNA imaging strategy were published online in the journal Nature Methods on April 6. In addition to Santangelo, Georgia Tech graduate student Aaron Lifland, Emory University associate professor Gary Bassell and Vanderbilt University professor James Crowe Jr. also contributed to this research. This research was funded by new faculty support from Georgia Tech.

In the study, the probes produced by attaching a few small fluorescent molecules called fluorophores to a modified nucleic acid sequence and combining the sequences with a protein exhibited single-molecule sensitivity and allowed the researchers to target and follow native RNA and non-engineered viral RNA in living cells.

"The great thing about these probes is that they recognize RNA sequences and bind to them using the same base pairing most people are familiar with in regards to DNA," explained Santangelo. "By adding only a few probes that would bind to a region of RNA, we gained the ability to distinguish a targeted RNA molecule from a single unbound probe because the former lit up two or three times brighter."

For their experiments, the team used a bacterial toxin to transport the probes into living cells a delivery technique that when combined with the high affinity of the probes for their targets, required significantly fewer probes than existing techniques. The toxin created several tiny holes in the cell membrane that allowed the probes to enter the cell's cytoplasm.

The researchers tested the sensitivity of conventional fluorescence microscopy to image individual probes inside a cell. Previous studies showed that these techniques were able to image an accumulation of probes inside a cell, but the current study demonstrated that individual probes without cellular targets could be observed homogenously distributed in the cytoplasm with no localization or aggregation.

With single-molecule sensitivity accomplished, the researchers investigated whether they could visualize individual RNA molecules using the probes. To do this, they simultaneously delivered probes designed to target a human messenger RNA (mRNA) sequence region and a probe designed with no target in the human genome. They were able to image unbound probes of both types as well as individual RNA molecules that had attached to the former probes.

The imaging technique also allowed the researchers to observe a process called dynamic RNA-protein co-localization, which is the joining of RNA molecules and RNA binding proteins in a single cell.

"We observed substantial transient interactions between proteins and viral RNA molecules that I don't think had ever been seen before with non-engineered RNA," noted Santangelo. "We saw one of the proteins move into a viral RNA granule and reside within it for over a minute before it was released, and we also saw another protein that appeared to dock with a viral RNA granule."

Santangelo is currently trying to improve the probes by making them smaller and brighter, while also using them to investigate viral pathogenesis and other biological phenomena.

"We are excited to use this imaging strategy to study how single viral RNAs travel from the nucleus of a cell to a virus assembly site, how mRNAs are regulated by location and time, and RNA trafficking in neurons," added Santangelo.


'/>"/>

Contact: Abby Vogel
avogel@gatech.edu
404-385-3364
Georgia Institute of Technology Research News
Source:Eurekalert  

Related biology technology :

1. Researchers peer into nanowires to measure dopant properties
2. Researchers find better way to manufacture fast computer chips
3. Collexis Connects Biomedical Researchers and Clinical Scientists with the Collexis Expert Platform for Translational Research
4. Researchers create catalysts for use in hydrogen storage materials
5. Thomson Reuters Names Hottest Researchers and Papers of 2007-2008
6. Researchers discover a potential on-off switch for nanoelectronics
7. University of Alberta and NINT researchers make solar energy breakthrough
8. Seminar Will Bring ALS Researchers to Patient Groups
9. Researchers at the University of Georgia Discover Protein is Crucial to Reproduction of Parasites Involved in Disease
10. FDA Approves Clinical Studies of a Novel Anti-Cancer Drug Developed by Italian Researchers
11. In Analyst Interview, Cord Blood America Says Stem Cells Now Front and Center for Researchers and Investors
Post Your Comments:
*Name:
*Comment:
*Email:
Related Image:
Researchers develop new way to see single RNA molecules inside living cells
(Date:2/8/2016)... 2016 Should antibiotic bone cement products be ... prevent infection after standard total hip or knee replacement ... Institute have been fielding a lot lately. ... Line?" --> "Antibiotic Bone Cement: ... --> While there isn,t a simple answer, ...
(Date:2/8/2016)... , February 8, 2016 ... Oncology Private Limited, an innovation-driven oncology company developing ... and less toxic, today announced that chairman emeritus ... has invested in the company as part of ... round, joining existing investors Navam Capital and Aarin ...
(Date:2/8/2016)... ... February 08, 2016 , ... Thomas ... is the Executive Vice President for Corporate Services and the Chief Financial Officer ... Payable, Payroll, Billing Operations, Treasury, Managed Care Contracting, Supply Chain, and Investments. , ...
(Date:2/8/2016)... Calverton, Maryland (PRWEB) , ... February 08, 2016 , ... ... our BSI Web product. The upgrade represents a completely new technical foundation and ... Engage will offer a responsive design interface, significantly increased speed for search results, a ...
Breaking Biology Technology:
(Date:2/4/2016)... , Feb. 4, 2016 The field ... one of the most popular hubs of the ... and other huge studies of human microbiota, have ... few years, the microbiome space has literally exploded ... research. This report focuses on biomedical aspects ...
(Date:2/2/2016)... 2016  BioMEMS devices deployed in hospitals ... medical screening and diagnostic applications, such as ... that facilitate and assure continuous monitoring without ... being bolstered through new opportunities offered by ... coupled with wireless connectivity and low power ...
(Date:2/2/2016)... 2, 2016  Based on its recent analysis ... recognizes US-based Intelligent Retinal Imaging Systems (IRIS) with ... for New Product Innovation. IRIS, a prominent cloud-based ... America , is poised to set the ... retinopathy market. The IRIS technology presents superior price-performance ...
Breaking Biology News(10 mins):