RNA

Part:BBa_K3218001:Design

Designed by: Abraham Grosman   Group: iGEM19_MSP-Maastricht   (2019-10-15)


Ribozyme - gRNA -Ribozyme cassette for expression of gRNA using self cleaveage mediated by the riboz


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 360
    Illegal NheI site found at 865
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 53
    Illegal NgoMIV site found at 82
    Illegal NgoMIV site found at 556
    Illegal NgoMIV site found at 585
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We had to consider the size of the plsmid if we were to include two gRNAs to be expressed, but this depends on if the dCas9-FokI system is being used which requires two gRNAs or the dCas9-AID which only requires one gRNA.


Source

The flanking ribozyme regions come from two separate genomes, the hammerhead ribozyme is present in several eukaryotic genomes and the Hepatitis delta virus ribozyme is derived from the hepatitis delta virus and are both known to self celave their phosphate backbone and are under the category of endonucleolytic ribozymes.


References