Composite

Part:BBa_K3168005

Designed by: Eva Hanckmann, Harm van der Veer, Claire Michielsen   Group: iGEM19_TU_Eindhoven   (2019-09-12)
Revision as of 13:55, 12 September 2019 by CMichielsen (Talk | contribs)


dCas9-SmallBitNanoLuc

This composite part is made up of two basic parts. The first basic part (CODE) codes for a catalytically dead CRISPR associated protein 9 (dCas9). This protein binds to a specific target on double stranded DNA. This specific target is determined by the guide RNA. The second basic part, which is fused to dCas9, consist of a (GGS)5 linker and the small bit of NanoLuc. When the small bit of NanoLuc forms a complex with the large bit, blue light is emitted. Thus this composite part is part of a Split-NanoLuc detection system, which targets a specific sequence on dsDNA and sends out a bioluminescent signal.

File:T--TU Eindhoven--dCas9-SmallBit.png

Figure 1. Schematic representation of dCas9-SmallBitNanoLuc.

Usage and Biology

dCas9 in combination with guide RNA forms a dsDNA recognition complex. A stronger bioluminescent signal is created when dCas9-LargBitNanoLuc and dCas9-SmallBitNanoLuc bind in close proximity.

T--TU Eindhoven--dCas9-SplitNL.png

Figure 2. Schematic representation of dCas9-Split-NanoLuc system.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1099
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3378
    Illegal BamHI site found at 4189
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None