Types | DnaRegion
|
Roles | Composite
engineered_region
|
Sequences | BBa_K1026010_sequence (Version 1)
|
Description
This gRNA targeting mRFP is a no-BioBrick-scars assembly of the red light regulated promoter BBa_K592006 (PFixK2), DNA sequence of a gRNA that targets mRFP reporter and a reliable terminator BBa_B0015. This gRNA sequence comes from the following literature: QI, LEI S., LARSON, MATTHEW H., GILBERT, LUKE A., DOUDNA, JENNIFER A., WEISSMAN, JONATHAN S., ARKIN, ADAM P. & LIM, WENDELL A. 2013. Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 152, 1173-1183.
Notes
gRNA shall be exactly transcribed out. Any extra nucleotides would potentially jeopardize the normal function of the gRNA. Therefore, BioBrick scars are not allowed here. Instead of cut-and-ligation, overlap PCR is preferred for the assembly task (changing promoter). In addition, a reliable terminator is applied here to ensure the termination of gRNA.
Source
The gRNA sequence is de novo synthesized.