dsRNA and RNAi dsRNA synthesis Technology of RNA Product
dsRNA synthesis
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 RNA 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 RNA will be intact (full-length) and free from any obvious RNase contamination. Although the RNA 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 RNA and analyzed on a 1 % agarose gel.
Experiment illustrating the consistency of dsRNA synthesis technology of RNA; 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. RNA 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.