Featured Research

from universities, journals, and other organizations

Rapid, accurate mRNA detection in plant tissues

Date:
April 17, 2014
Source:
American Journal of Botany
Summary:
Messenger RNA (mRNA) plays an important role in gene expression, and examining the types and amounts of mRNA present in an organism allows researchers to answer key questions about gene expression and regulation. A recent study shows that RNAScope ISH (developed for studies in animal -- particularly human -- tissues) is faster and more sensitive than traditional ISH in detecting and quantifying mRNA in plants.

Gene expression is the process whereby the genetic information of DNA is used to manufacture functional products, such as proteins, which have numerous different functions in living organisms. Messenger RNA (mRNA) serves as an important intermediary during gene expression, by relating the genetic information of DNA to the molecular mechanisms involved in manufacturing proteins.

By examining the different types and amounts of mRNA molecules present in an organism at a given time, researchers can determine which specific genes are being expressed. This, in turn, offers tremendous insight into the genes responsible for producing different morphological forms (or phenotypes).

Scientists generally rely on in situ hybridization (ISH) techniques (using mRNA-specific probes) to determine the presence or absence of particular mRNA molecules in plant tissues. Most traditional ISH methods, however, are time- and labor-intensive and lack the sensitivity necessary to precisely quantify the amount of expression of each gene.

In a new study in the April issue of Applications in Plant Sciences, researchers from Dow AgroSciences demonstrate the effectiveness of a new ISH method, called RNAscope ISH, for studies of gene expression in plants. In contrast to traditional approaches, RNAScope ISH is significantly faster and highly sensitive, permitting researchers to not only detect but also quantify, with confidence, the expression levels shown by genes of interest.

RNAScope ISH was developed by Advanced Cell Diagnostics (ACD) Inc., initially for studies of gene expression in animal (and especially human) tissues. It is a type of branched DNA ISH that uses pairs of 'Z-probes,' which are highly specific to target genes, but also small enough to easily diffuse into the tissues under study. Once the Z-probes bind to the mRNA molecules of interest, this allows label molecules to also bind, which then makes the mRNA molecules detectable, either via fluorescence or chromogenic detection.

According to Dr. Andrew Bowling, lead author of the paper and a research scientist at Dow Agrosciences, detected mRNA molecules appear as individual spots within the cell, making the expression results very easy to interpret.

"This method has a very low background level, which allows for higher confidence in the results obtained. With the conventional method, there seemed to always be a point where you were trying to determine if one tissue or cell was 'bluer' than another, or 'bluer' than the control. With the RNAscope method, the experimental sections have spots and the control sections don't. The results are very clear."

Bowling adds that RNAscope ISH is also a very accessible method compared to more traditional ISH approaches.

"The vendor provides probe design and synthesis services, greatly reducing the suite of skills that are required to carry out a successful ISH experiment. You don't have to be (or collaborate with) an RNA in vitro transcription expert to make your labeled probes. You don't have to synthesize plasmids to transcribe. You don't have to run northern blots to confirm efficacy and specificity. That is all part of the probe design and synthesis service. Also, it only takes about two weeks from emailing them a sequence to obtaining the probe in the mail from them."

"Furthermore, the [RNAScope ISH] method is faster. It takes about 8-10 hours to do an experiment. The classic ISH methods could sometimes require one or even two overnight incubations."


Story Source:

The above story is based on materials provided by American Journal of Botany. Note: Materials may be edited for content and length.


Journal Reference:

  1. Andrew J. Bowling, Heather E. Pence, Jeffrey B. Church. Application of a Novel and Automated Branched DNA in Situ Hybridization Method for the Rapid and Sensitive Localization of mRNA Molecules in Plant Tissues. Applications in Plant Sciences, 2014; 2 (4): 1400011 DOI: 10.3732/apps.1400011

Cite This Page:

American Journal of Botany. "Rapid, accurate mRNA detection in plant tissues." ScienceDaily. ScienceDaily, 17 April 2014. <www.sciencedaily.com/releases/2014/04/140417090836.htm>.
American Journal of Botany. (2014, April 17). Rapid, accurate mRNA detection in plant tissues. ScienceDaily. Retrieved September 30, 2014 from www.sciencedaily.com/releases/2014/04/140417090836.htm
American Journal of Botany. "Rapid, accurate mRNA detection in plant tissues." ScienceDaily. www.sciencedaily.com/releases/2014/04/140417090836.htm (accessed September 30, 2014).

Share This



More Plants & Animals News

Tuesday, September 30, 2014

Featured Research

from universities, journals, and other organizations


Featured Videos

from AP, Reuters, AFP, and other news services

Earth Has Lost Half Its Vertebrate Wildlife Since 1970: WWF

Earth Has Lost Half Its Vertebrate Wildlife Since 1970: WWF

Newsy (Sep. 30, 2014) A new study published by the World Wide Fund for Nature found that more than half of the world's wildlife population has declined since 1970. Video provided by Newsy
Powered by NewsLook.com
Dolphins Might Use Earth's Magnetic Field As A GPS

Dolphins Might Use Earth's Magnetic Field As A GPS

Newsy (Sep. 30, 2014) A study released Monday suggests dolphins might be able to sense the Earth's magnetic field and possibly use it as a means of navigation. Video provided by Newsy
Powered by NewsLook.com
How To Battle Stink Bug Season

How To Battle Stink Bug Season

Newsy (Sep. 30, 2014) Homeowners in 33 states grapple with stink bugs moving indoors at this time of year. Here are a few tips to avoid stink bug infestations. Video provided by Newsy
Powered by NewsLook.com
California University Designs Sustainable Winery

California University Designs Sustainable Winery

Reuters - US Online Video (Sep. 27, 2014) Amid California's worst drought in decades, scientists at UC Davis design a sustainable winery that includes a water recycling system. Vanessa Johnston reports. Video provided by Reuters
Powered by NewsLook.com

Search ScienceDaily

Number of stories in archives: 140,361

Find with keyword(s):
Enter a keyword or phrase to search ScienceDaily for related topics and research stories.

Save/Print:
Share:

Breaking News:

Strange & Offbeat Stories


Plants & Animals

Earth & Climate

Fossils & Ruins

In Other News

... from NewsDaily.com

Science News

Health News

Environment News

Technology News



Save/Print:
Share:

Free Subscriptions


Get the latest science news with ScienceDaily's free email newsletters, updated daily and weekly. Or view hourly updated newsfeeds in your RSS reader:

Get Social & Mobile


Keep up to date with the latest news from ScienceDaily via social networks and mobile apps:

Have Feedback?


Tell us what you think of ScienceDaily -- we welcome both positive and negative comments. Have any problems using the site? Questions?
Mobile: iPhone Android Web
Follow: Facebook Twitter Google+
Subscribe: RSS Feeds Email Newsletters
Latest Headlines Health & Medicine Mind & Brain Space & Time Matter & Energy Computers & Math Plants & Animals Earth & Climate Fossils & Ruins