Advantage and Core Benefit
- Auxin-Inducible Degron (AID) protein inhibition is much faster than siRNA/antisense.
- Improved sensitivity, reduced cytotoxicity and applicable for various cell types.
- Single step preparation for in-vitro experiment by CRISPR/Cas9 technology.
Background and Technology
Conditional gene knockout/down (KO/KD) is widely used for drug discovery as well as basic bioscience research. Among several methods; siRNA/antisense KD (1), Cre/loxP system (2), targeted protein degradation (3), etc., 1 and 2 are commonly used, but takes long time before natural degradation of target protein. Targeted protein degradation (3) has an advantage because it can rapidly inhibit the function by degrading cytoplasmic/membrane/nuclear protein in an hour or so. Several methods of 3 reported have pros and cons. AID is a simple method, but high concentration auxin is required, that causes cytotoxicity.
We have modified auxin and its receptor TIR1, and identified a pair that dramatically reduces effective concentration to nM order (Fig).
Pat. pending (not published)
Journal published; DOI:10.1093/nar/gkaa748
Kohei Nishimura (Nagoya University), et al.
We will grant license to companies who have CRISPR/Cas9 license.
We are also seeking companies who develop conditional AID animal, in corporation with us.
Modified TIR1 vector can be provided under Material transfer agreement.
Auxin derivative 5-Ad-IAA is commercially available.
Product No. KJ-03248