Part:BBa_K5047031
DNA encodes a shRNA binds to chitin synthase gene of V. velutina.
This DNA sequence encodes a short hairpin RNA shRNA designed to target a specific region within the chitin synthase gene of the Asian hornet Vespa velutina. The target region is located on an exon common to all isoforms of the chitin synthase gene GeneID 124951980 Genbank XM_047501172.1 within the genomic coordinates NC_062188.1 19915311-19930067 iVesVel2.1 genome GCF_912470025.1.
The specific target sequence spans from positions 19925853 to 19925878. This shRNA has been designed to keep GC content intermediate approx. 30 - 50% with 48% in this variant to avoid strong secondary structure. Additionally, this shRNA is the variant with the lowest number of potential off-target interactions compared to TS2 BBa_K5047022 TS3 BBa_K5047025 and TS4 BBa_K5047028. This sequence has off-targets in Vespa mandarinia and Vespa crabro with one mismatch only.
Upon testing in human embryonic kidney 293T cells using a luciferase assay, Schopman Nick C T et al. “Optimization of shRNA inhibitors by variation of the terminal loop sequence.” Antiviral Research vol. 86.2 (2010): 204-11. doi: 10.1016/j.antiviral.2010.02.320 reported that shRNAs based on the mir-17 and mir-25 loops exhibited the strongest target repression, approximately 10-fold stronger than the 9-nt loop.
This shRNA contains the optimized UGAUAUGUGCA loop Schopman Nick C T et al. Optimization of shRNA inhibitors by variation of the terminal loop sequence Antiviral research vol 86.2 2010 204-11 doi: 10.1016/j.antiviral.2010.02.320. Through secondary structure analysis performed by ViennaRNA the free energy of the thermodynamic ensemble is -50.39 kcalmol. The frequency of the minimum free energy structure in the ensemble is 52.98%. The ensemble diversity is 0.70.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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