Part:BBa_K3168005
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.
Figure 2. Schematic representation of dCas9-Split-NanoLuc system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1099
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3378
Illegal BamHI site found at 4189 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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