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About Us

IVD Medical Instrument Business

Nucleic Acid Extracting Instrument

Instrument which makes it possible to extract nucleic acid quickly and efficiently from various and plenty of samples.

Nucleic Acid Extraction Reagents

Reagents which facilitate extraction of DNA/RNA from samples collected from human body (Urine, Swab, Sputum, Blood, Plasma, Serum, etc.).

Other Molecular Diagnostics Products

Reagents for extracting DNA/RNA of pathogenic using Real Time PCR.

RNAi Business

RNAi Service (siRNA/shRNA synthesis and other service)

siRNA/shRNA synthesis service to inhibit gene expression and other gene-related service.

Long dsRNA Synthesis Service

Long dsRNA synthesis to inhibit gene expression by injecting RNA into various living organisms (Inhibition of gene expression for development of herbicide/insecticide).

dsRNA and RNAi

RNA interference(RNAi) is a novel method of sequence-specifically blocking gene function that works by introducing into cells with short sequences of RNA that match part of a target organism's gene sequence.

RNA interference works in the cytoplasm of the cell; thus it does not genetically modify the organism. It can block an organism's production of an essential protein, suppress or modify various genetic traits in an organism without changing the genetic make-up of the organism. It has been well known that long dsRNA (200-800bp) can initiate RNAi in plant, worms, insect, and even shrimp. Thus, dsRNA may be a new raw material for herbicide, insecticide, anti-fungal and anti-viral applications for agricultural as well as environmental applications. Because of its sequence specificity, it can also be designed to have no toxicity to unintended organisms.

An RNA approach, especially when using long dsRNAs, can also minimize the development of insecticidal or herbicidal resistance, and be a key objective in the development of agricultural control agents.

In contrast to the difficulty of delivering RNA in humans and animals, an RNAi gene silencing effect by dsRNA has been shown in plants, insects, worms, and crustaceans by a simple spray, injection, or feeding.

Even just spraying crude extracts of bacterially expressed dsRNA or in vitro transcribed dsRNA can be used to protect plants against several viral infections.

Also a dsRNA corresponding to an essential insect gene is sprayed on a crop. As the insect is feeding on the crop, it takes up the dsRNA which in turn eliminates or reduces an essential protein in the insect leading to the death of the insect and protection of the crop plant. Another approach might be to make a dsRNA corresponding to a herbicide-resistance gene in weeds thus rendering the weed susceptible to the herbicide. dsRNAs have thus emerged as an innovative pest and weed control reagent.

dsRNA Synthesis and Purification

Chemical synthesis of long dsRNA is not feasible for agricultural applications due to the limit of technology and the high cost.
There are two non-chemical sources of dsRNA: in vivo synthesis and in vitro transcription. There are Pros and Cons for each method as described in the next Table.

Method	time and labor demand	Cost/unit RNA	Biosafety	Purification	Mass production
In vivo	High	HighAlso requires a large fermenting system	Considerable concern	Hard	Possible
In vitro	Medium	Moderate	N/A	Easy	Maybe

For field application, dsRNA should be made according to the following three rules:

First, dsRNA should be supplied in bulk in a cost-effective manner.
Second, the RNA does not need to be as pure as for medical application.
Third, the source of the dsRNA should entail minimal GMO concerns. Considering all of these issues, AgroRNA has developed the most effective procedures for the synthesis of dsRNA of mg to kg scale ranging from 200 to 800 bp in a manner which is free from any GMO or bio-safety issues.
The technology is based on the most effective method of dsRNA synthesis per supplied rNTP unit developed by AgroRNA.

The initial goal of AgroRNA was just the bulk production of dsRNA in a practical price for the agricultural application.

Additional interests are also being focused into the purification of dsRNA from the reaction mixture for the regulation approval issue. Development of the purification method of dsRNA from the bulk transcriptional reaction is essential. The final products should be free from the major contaminants of truncated dsRNA, free rNTP, enzymes, salts, and DNA template. Also, the method should meet all feasibility issues of cost, efficacy, and also for the bulk volume handling.

AgroRNA has been developed the method for dsRNA purification from the bulk reaction mixture without the major contaminants in cost effective manner. As shown in Figure 1 , the purified sample contains more dsRNA with less contaminants such as free rNTP.

Figure1. The amount of dsRNA after each step of purification. Total 1 liter of reaction mixture for a 450 bp dsRNA was further processed for purification. The amount of RNA was determined by Nanodrop after appropriated dilution. Total 1 ug of each RNA was loaded on the 1% of Agarose gel after each step of purification. Lane 1; before any purification, lane 2; after the 1st step of purification, lane 3; after 2nd step of purification, and lane 4; after 3rd step of purification. Even with the same OD values on Nanodrop, the total amount of dsRNA is dramatically increased after final steps of purification.

Most strikingly, we have found that the efficacy of dsRNA for gene silencing to a target insect is drastically increased using the purified RNA as shown Figure.

The data clearly indicates the purification of dsRNA is essential for better efficacy in bioassay. AgroRNA will supply dsRNA without or with appropriated purification process to meet the diverse demands from multiple customers.

Figure2. The dsRNA mediated motility to a target insect is highly dependent on the purity of the dsRNA. Total 20 of each target insect was fed with plant sprayed with dsRNA of control (GFP dsRNA) or test dsRNA (target to b-Tubulin) before (left panel) or after purification (right panel), respectively. The survival of insect was counted in each indicated hour. The data is average of three independent trials.

Technology of AgroRNA

dsRNA synthesis and purification :

Although an initial proof-of-concept for dsRNA-mediated RNAi in plant and insect has been shown, commercial application is highly depended on the supply of bulk amount of RNA at price points compatible with such applications and competitive with traditional chemical reagents.

Even for the construction of a GMO plant, the target gene screening is time and labor intensive unless there was a prompt and convenient supply of dsRNA.

For the effective, but massive production of dsRNA, the agroRNA developed several proprietary procedures and methods for dsRNA synthesis, purification, and preservation. The procedures include a bulk NTP synthesis and purification, bulk enzyme preparation and purification, enzyme recycling system, bulk transcription system, and novel dsRNA purification system.

All dsRNA synthesized by the agroRNA will be intact (full-length) and free from any obvious RNase contamination. Although the agroRNA can make more than 2 Kb dsRNA, we will supply 100-800 bp dsRNA products for optimum RNAi activity and cost effective production. The figure shows several dsRNA products synthesized by agroRNA and analyzed on a 1 % agarose gel.

Experiment illustrating the consistency of dsRNA synthesis technology of agroRNA; two different dsRNA products were synthesized in three independent trials and analyzed on a 6% PAGE gel.

Target discovery and formulation :

Key to success in plant RNAi against agriculturally important pest is not only the identification of suitable gene targets, but also the expression and delivery of sufficient amount of intact dsRNA to the pest organism. There are several agriculturally important pests such as cotton bollworm, corn rootworm, potato beetle, and mites. RNAi directed against a number of gene targets such as tubulin, or a group of ATPases was effective in several economically important pests by simple feeding of the dsRNA.

To apply dsRNA as a potential insecticide or herbicide with high specificity and efficacy, screening for target genes and its optimal dsRNA are essential steps. By introducing each dsRNA into the target organism, the optimum target gene and its corresponding target dsRNA sequence can be determined empirically. Based on the phenotype being screened for, the most effective target gene and its corresponding dsRNA sequence can be determined. agroRNA is screening target genes using multiple dsRNA sequences against several economically/agriculturally important pests. After the target gene discovery, dsRNA will be tested in small or medium field tests as sprayable pesticides. Alternatively the dsRNA may be further formulated to enhance delivery or introduced as a transgene to create a pest resistant corp.

Products

All dsRNAs supplied by agroRNA have the following features.

Proprietary dsRNA synthesis platform enables mutation-free dsRNA of 200 to 800 bp.
The sequence of dsRNA may be from any organism and any sequence.
All synthesized dsRNA sequence information will be regarded as confidential.
The information will not be released to others under any circumstances.
The products are completely free from any living organisms such as genetically modified E. coli with expresses dsRNA.
The product will be supplied with analysis data by an Agarose gel and Nanodrop.
The quality and exact size of dsRNA product can be assured.
The sequence of supplied RNA is confirmed by sequencing.
All supplied dsRNA will be pre-annealed as a duplex.
The supplied dsRNA will not contain apparent amounts of truncated or degraded products which could reflect RNase contamination.
dsRNA will be supplied without or with purification as shown in next section.
RNA will be shipped at ambient temperature and can be stored at 4oC (for less than a month) or -20oC (for more than a month) upon receiving.

dsRNA grades from AgroRNA

*table

Grade	Features	Included components	Purity	Scale
Grade1	Synthesis mixture without any purification.	Major dsRNA Major free NTP Enzymes and salt	50% or less	To 1 Kg
Grade2	Supply in DW	Major dsRNA only Without size selection	Up to 95%	To 1 Kg
Grade3	HPLC grade Supply in DW	Major dsRNA only After a partial size selection	Up to 95%	To 100gram scale

Grade

Features

Included components

Purity

Scale

Grade1

Synthesis mixture without any purification.

Major dsRNA

Major free NTP

Enzymes and salt

50% or less

To 1 Kg

Grade2

Supply in DW

Major dsRNA only

Without size selection

Up to 95%

To 1 Kg

Grade3

HPLC grade

Supply in DW

Major dsRNA only

After a partial size selection

Up to 95%

To 100gram scale

Remarks :

Shipping at ambient temperature and store at 4oC (for less than a month) or -20oC (for more than a month).
Supplied amount is about 70% of order scale. (effective on January 1, 2018)
There is variation for the supplied amount depends on the dsRNA size. Typically, less amount for shorter dsRNA.

dsRNA Storage

The RNA in the shipping and storage solution can be stored at ambient temperature for more than 3 months without any sign of degradation. The RNA can be further stored in -20oC for an indefinite period.

dsRNA of 839 base in the storage solution was stored at 37oC incubator.

The RNA was analyzed on 1 % agarose gel in the indicated day.

Please Fill out the attached order form (link) and send to AgroRNA through email (info@agrorna.com) with a formal PO number and your Fedex account number.
AgroRNA will deliver your dsRNA in 14-20 (less than 10g) or 30 days (more than 10g) after the order.
The supplied dsRNA will be delivered with analysis data of agarose gel and Nanodrop.
Based on the data, the exact size and purity of dsRNA product can be assured.
Without supply of your Fedex account number, we will charge and add the appropriate shipping cost to your invoice.

Synthesis	Guaranteed amount of dsRNA*	Turn-around time
10mg scale	More than 70%	2-3 weeks
100mg scale	More than 70%	2-3 weeks
1g scale	More than 70%	2-3 weeks
10g scale	More than 70%	2-3 weeks
100g scale	More than 50%	1 month
1Kg scale	More than 50%	1~2 month

Special Partnership Program

dsRNA Company has established a special strategic partnership program for dsRNA synthesis service to clients worldwide.
In particular, this new program offers the service at special and competitive prices to our valued clients who want to establish a long-term preferred vendor contract with dsRNA Company.
We are willing to work with you in a flexible manner within your budget to achieve your goals. With our solid growth and financial performance, dsRNA Company assures reliable and sustainable services.