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Kanemaki’s Auxin Inducible Degron (AID) System


To investigate the role of proteins in cells, gene knock down method is widely applied. Here Dr. Kanemaki offers a new method of reduction of interested proteins. He provides a novel method Auxin Inducible Degron (AID) System with which you can degrade proteins fused with a degron (a small domain that can induce protein degradation) in a very short period of time.

DNA Resource

Natsume, T., Kiyomitsu, T., Saga, Y., Kanemaki, M.T. Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors. Cell Reports 15: 210-218, 2016. PMID: 27052166.

Table. Available fluorescent and selection markers.

Selection
marker
Attached tag at the C-terminus
mAID mAID-mClover mAID-mCherry2
Neo pMK286 pMK289 pMK292
Hygro pMK287 pMK290 pMK293
Bsr pMK288 pMK291 pMK294

 
Table 2. CsTIR1 expression vectors and helper plasmid to knock-in the expression vectors.

Catalog# Name of clone Running title
RDB13918 pMK232 OsTIR1 expression vector
RDB13919 pMK243 OsTIR1 expression vector
RDB13917 AAVS1 T2 CRIPR in pX330 Helper for knock-in the expression vector to the human AAVS1 site

 
Table 3. Tags without mAID are also available.

Selection
marker
Attached tag at the C-terminus
mClover mCherry2 S tag-3FLAG
Neo pMK277 pMK280 pMK283
Hygro pMK278 pMK281 pMK284
Bsr pMK279 pMK282 pMK285

Please refer Fig. 4 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use these clones.


Nishimura, K., Fukagawa, T., Takisawa, H., Kakimoto, T., Kanemaki, M. An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nat. Methods., 6, 917-922, 2009. PMID: 19915560.

Catalog# Name of clone Running title Type
RDB08468 pNHK60 Expression clone for an auxin-based degron (AID) system Plasmid
RDB08469 pMK106 Expression clone for an auxin-based degron (AID) system Plasmid
RDB08470 pMK107 Expression clone for an auxin-based degron (AID) system Plasmid

Please refer
Supplementary Figure 11 for pMK106 (aid-CENPH)
Supplementary Figure 11 for pMK107 (CENPH-aid)
Supplementary figure 10 for pNHK60 (pIRES OsTIR1-EGFP-IAA17)

map of RDB08468

 


 

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DNA resources containing GFP and mutants such as CFP and YFP&nbdp;[link]
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References & Related Articles

  1. Natsume, T., Kiyomitsu, T., Saga, Y., Kanemaki, M.T. Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors. Cell Reports, in press, Published online: March 24, 2016. doi: 10.1016/j.celrep.2016.03.001.
  2. Nishimura, K., Fukagawa, T., Takisawa, H., Kakimoto, T., Kanemaki, M. An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nat. Methods., 6, 917-922, 2009. PMID: 19915560.
  3. Kanemaki, M.T. Frontiers of protein expression control with conditional degrons. Eur. J. Physiol., 465, 419-425, 2013. PMID: 23271452.
  4. Matato Kanemaki Lab Web Page
  5. BioResource Now! Issue 7, July 2011 (PDF, in Japanese)
  6. BioResource Now! Issue 7, July 2011 (PDF, in English)

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2017.11.30