Abstract
Background. The perspective of research of small interfering RNA (siRNA) application in medical practice was proved by its efficiency of chemically synthesized siRNA in the experiment in vitro . The high cost of RNA production through chemical synthesis determines the importance of searching the biotechnological methods of obtaining these compounds. A promising area of biopharmaceuticals creation is a combination of active pharmaceutical substances with innovative tools of delivery, such as bio-encapsulation in plant cells. Objective. The aim of the paper is testing of capabilities and efficiency of transient expression of the gene encoding the small interfering RNA to mRNA \[\delta-isoform\] of human protein kinase C \[(anti-PKC\delta)\] in edible raw plant biomass of lettuce Lactuca sativa . Methods. Obtaining the biomass which accumulates anti-PKCd small interfering RNA molecules was carried out by the method that was developed by our research group for the expression of genes encoding recombinant proteins. Evaluation of small interfering RNA accumulation was performed by a reverse transcription method with the subsequent Real-Time Polymerase Chain Reaction (PCR). Results. The level of accumulation of target product is 22 fmol/g of lyophilized plant biomass. Conclusions. The method of transient expression allows obtaining the lettuce Lactuca sativa biomass, which contains small interfering RNA recombinant anti-PKCd (detected by a reverse transcription method with the subsequent Real-Time PCR).
Highlights
The perspective of research of small interfering RNA application in medical practice was proved by its efficiency of chemically synthesized siRNA in the experiment in vitro
A novel chemopreventive strategy based on therapeutic microRNAs produced in plants / S
Oral delivery of protein drugs bioencapsulated in plant cells // Molecular Therapy. — 2016. — 24. — No 8. — P. 1342—1350
Summary
The perspective of research of small interfering RNA (siRNA) application in medical practice was proved by its efficiency of chemically synthesized siRNA in the experiment in vitro. Obtaining the biomass which accumulates anti-PKCd small interfering RNA molecules was carried out by the method that was developed by our research group for the expression of genes encoding recombinant proteins. The method of transient expression allows obtaining the lettuce Lactuca sativa biomass, which contains small interfering RNA recombinant anti-PKCd (detected by a reverse transcription method with the subsequent RealTime PCR). Що, за аналогією в різниці рівнів накопичення рекомбінантних білків, саме транзієнтна експресія генів малих інтерферуючих РНК у рослинах потенційно теж має дати можливість отримувати вищі рівні накопичення цих молекул, ніж експресія вбудованих у геном рослини трансгенів
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