PMID- 21397955
OWN - NLM
STAT- MEDLINE
DCOM- 20110729
LR  - 20110408
IS  - 1873-2534 (Electronic)
IS  - 0165-2427 (Linking)
VI  - 141
IP  - 1-2
DP  - 2011 May 15
TI  - Targeted knockdown of canine KIT (stem cell factor receptor) using RNA 
      interference.
PG  - 151-6
LID - 10.1016/j.vetimm.2011.02.006 [doi]
AB  - Canine mast cell tumours often express KIT mutations that result in constitutive 
      activation of the c-kit receptor and which are associated with more aggressive 
      disease. The aim of the current study was to determine whether small inhibitory 
      RNA (SiRNA) molecules could specifically target canine KIT mRNA for knock-down. 
      Canine beta-2 microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase 
      (GAPDH) and KIT sequences were cloned into the psiCHECK-2 vector. SiRNA 
      molecules, designed to target gene-specific sequences, were co-transfected with 
      plasmid DNA into Chinese hamster ovary (CHO) cells. Renilla and firefly 
      luciferase activity was measured using the Dual-GLO((R)) Luciferase Assay 
      (Promega). Using this reporter system, canine housekeeping gene-specific SiRNA 
      molecules demonstrated knockdown of their targets (72.0% knockdown for B2M and 
      94.5% knockdown for GAPDH). An SiRNA molecule targeting exon 2 of canine KIT 
      successfully knocked-down reporter gene expression of a KIT(26-407) construct 
      (90.8% knockdown). An SiRNA molecule targeting a 48 base-pair in-tandem 
      duplication mutation in KIT exon 11 selectively knocked down expression of the 
      KIT(1569-1966mutant) construct (93.1% knockdown) but had no effect on the 
      KIT(1569-1918wild-type) construct. The results show that RNA interference can be 
      used to inhibit canine KIT mRNA expression and has the potential to selectively 
      target the mutant version of KIT that is expressed by some malignant mast cells.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Elders, R C
AU  - Elders RC
AD  - Department of Veterinary Clinical Sciences, Royal Veterinary College, Hawkshead 
      Lane, North Mymms, Hatfield, Hertfordshire AL97TA, UK.
FAU - Holder, A
AU  - Holder A
FAU - Baines, S J
AU  - Baines SJ
FAU - Argyle, D
AU  - Argyle D
FAU - Catchpole, B
AU  - Catchpole B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110223
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (RNA, Small Interfering)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-kit)
RN  - EC 3.1.1.3 (CEL protein, human)
RN  - EC 3.1.1.3 (Lipase)
SB  - IM
MH  - Animals
MH  - Dogs/*genetics
MH  - Gene Expression Regulation/genetics
MH  - Gene Knockdown Techniques/methods/*veterinary
MH  - Lipase
MH  - Proto-Oncogene Proteins c-kit/*genetics
MH  - *RNA Interference
MH  - RNA, Small Interfering/genetics
MH  - Transfection/veterinary
EDAT- 2011/03/15 06:00
MHDA- 2011/07/30 06:00
CRDT- 2011/03/15 06:00
PHST- 2010/05/14 00:00 [received]
PHST- 2011/01/25 00:00 [revised]
PHST- 2011/02/14 00:00 [accepted]
PHST- 2011/03/15 06:00 [entrez]
PHST- 2011/03/15 06:00 [pubmed]
PHST- 2011/07/30 06:00 [medline]
AID - S0165-2427(11)00049-3 [pii]
AID - 10.1016/j.vetimm.2011.02.006 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2011 May 15;141(1-2):151-6. doi: 
      10.1016/j.vetimm.2011.02.006. Epub 2011 Feb 23.