SARS-CoV-2 belongs to the genus of beta-coronaviruses and causes a novel type of severe acute respiratory syndrome known as COVID-19. The 30-kb genome consists of a positive-sense single-stranded RNA (+ssRNA), which encodes a set of non-structural and structural proteins (1). Amongst the latter, the spike surface glycoprotein (S-protein) is responsible for the human host-receptor binding, which was identified as the angiotensin-converting enzyme 2 (ACE2) (2).
With the sudden appearance of SARS-CoV-2 in December 2019 mechanisms on the host-virus interaction and the cellular replication are still largely unknown. But the vast global spread requires fast and efficient research solutions to find potential treatments and vaccines.
Endoribonuclease-prepared siRNA as SARS-CoV-2 research tool
Eupheria Biotech developed the esiRNA technology (endoribonuclease-prepared siRNAs) that provides a highly specific knockdown of transcripts by RNA interference (RNAi). With the longstanding expertise in the functional characterization of genes, Eupheria adapted the esiRNA technology to provide customers with a tool for fast and reliable SARS-CoV-2 research.
Based on recent publications (2, 3, 4, and others) we designed an endoribonuclease-prepared siRNA library that covers more than 350 human target genes. These include human interaction partners of SARS-CoV-2 proteins, known or potential host-virus factors of the COVID-19 and other coronaviruses, such as SARS-CoV and MERS, genes involved in intracellular virus progression and druggable targets for the identification of potential treatments.
We offer this library as a fully customized esiFLEX option in a 96-well plate format at scales of 5.0 µg sufficient for screening diverse human cell lines. Additional genes can be added at a free choice and single targets can be ordered as individual esiRNA. Contact our technical support for information on all targets of the panel at info©eupheria.com.
Our COVID-19 esiRNA library is the perfect research tool for fast and easy screening and rapid identification of host-virus factors. For more information visit our esiRNA product page and see our Tools & Resource Section “How to handle your esiRNA” for recommendations concerning the transfection conditions in different cell lines.
(1) Wu et al., Genome Composition and Divergence of the Novel Coronavirus (2019-nCoV) Originating in China. Cell Host & Microbe 27 (3), March 2020, pp 325-328
(2) Hoffmann et al., SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and is Blocked by a Clinically Proven Protease Inhibitor. Cell 181 (2), April 2020, pp 271-280.e8
(3) Li et al., Assessing ACE2 expression patterns in lung tissue in the pathogenesis of COVID-19. Journal of Autoimmunity 112, August 2020
(4) Tsimbas et al., Coronaviruses in Hematologic Malignancies: Targeting Molecules Beyond the Angiotensin-Converting Enzyme 2 (ACE2) Wall in COVID-19. Pathology and Oncology Research, April 2020
All esiRNAs were designed based on the sequence annotations from the ENSEMBL database.
Quality Control: esiRNAs are designed using the DEQOR design algorithm. The PCR products used for production are tested via gel electrophoresis and identity is confirmed by DNA sequencing. Quality of esiRNA is validated by gel electrophoresis and concentrations are determined by measuring OD260 or fluorescence intensity.
Purification: RNaseIII-digested products are purified via column purification using anion-exchange chromatography, precipitated with isopropanol and washed with ethanol.
Format: All esiRNA are delivered in solution (resuspended in nuclease-free TE buffer) normalized to 50 ng/µl.
Storage and Stability: esiRNAs stored at -20°C are stable for 2 years. No decrease in stability was observed after 12 freeze/thaw cycles.
Shipment: Shipped in solution on dry ice in sealed 96-well plates.
Technical Data Sheet: The COVID-19 panel will be delivered with a CD containing the technical data sheet, which includes the plate number and positions of esiRNAs, the esiRNA ID numbers, organism, amount (µg), concentration, lot #, and target sequence.