Difference between revisions of "Part:BBa K5047003"
Line 3: Line 3:
  
  
−
<!-- This DNA sequence encodes a shRNA which can be used as a control in RNA interference experiments. Instead of targeting a functional gene this sequence contains two inverted repeats of a sgRNA derived from a CRISPR interference system separated by a 9nt loop Brummelkamp Thijn R et al A system for stable expression of short interfering RNAs in mammalian cells Science New York NY vol 2965567 2002 550-3 doi1061126science1068999. The shRNA does not have any significant biological target in our model organism making it an ideal negative control to verify the specificity and functionality of the RNA interference RNAi machinery in our experiments. This ensures that any observed effects in the system are due to the specific shRNAs being tested and not due to nonspecific or off-target interactions.
+
<!-- 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_062188.1:19,915,311-19,930,067 (iVesVel2.1 genome, GCF_912470025.1). The specific target sequence spans from positions 19’925’853 to 19’925’878. 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_XXXXX), TS3 (BBa_XXXXX) and TS4 (BBa_XXXXX). This sequence has off-targets in Vespa mandarinia and Vespa crabro with one mismatch only.
  
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 12:28, 30 September 2024

DNA sequence encodes a shRNA that does bind to the chitin synthase gene of V. velutina.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]