Difference between revisions of "Part:BBa K3168005"

Revision as of 12:23, 23 September 2019

dCas9-SmallBitNanoLuc

This composite part is made up of two basic parts (figure 1). The first basic part (BBa_K3168000) codes for a catalytically dead CRISPR associated protein 9 (dCas9). dCas9 binds to a specific double-stranded DNA sequence which is determined by the guide RNA. The second basic part (BBa_K3168003), which is fused to dCas9, consists 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 upon binding of this specific sequence (figure 2).

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-LargeBitNanoLuc and dCas9-SmallBitNanoLuc bind in close proximity.

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

Sequence and Features