PMID- 27142766
OWN - NLM
STAT- MEDLINE
DCOM- 20170302
LR  - 20181113
IS  - 1543-706X (Electronic)
IS  - 1071-2690 (Linking)
VI  - 52
IP  - 7
DP  - 2016 Aug
TI  - Complete reduction of p53 expression by RNA interference following heterozygous 
      knockout in porcine fibroblasts.
PG  - 736-41
LID - 10.1007/s11626-016-0026-0 [doi]
AB  - Tumor suppressor p53 plays a critical role in the regulation of cell cycle and 
      apoptosis in mammals. Mutations of p53 often cause various cancers. Murine models 
      have improved our understanding on tumorigenesis associated with p53 mutations. 
      However, mice and humans are different in many ways. For example, the short 
      lifespans of mice limit the clinical application of the data obtained from this 
      species. Porcine model could be an alternative as pigs share many anatomical and 
      physiological similarities with humans. Here, we modified the expression levels 
      of p53 messenger RNA (mRNA) and protein in porcine fetal fibroblasts using a 
      combination of gene targeting and RNA interference. First, we disrupted the p53 
      gene to produce p53 knockout (KO) cells. Second, the p53 shRNA expression vector 
      was introduced into fibroblasts to isolate p53 knockdown (KD) cells. We obtained 
      p53 KO, KD, and KO + KD fibroblasts which involve p53 KO and KD either separately 
      or simultaneously. The mRNA expression of p53 in p53 KO fibroblasts was similar 
      to that in the wild-type control. However, the mRNA expression levels of p53 in 
      KD and KO + KD cells were significantly decreased. The p53 protein level 
      significant reduced in p53 KD. Interestingly, no p53 protein was detected in KO + 
      KD, suggesting a complete reduction of the protein by synergistic effect of KO 
      and KD. This study demonstrated that various expression levels of p53 in porcine 
      fibroblasts could be achieved by gene targeting and RNA interference. Moreover, 
      complete abolishment of protein expression is feasible using a combination of 
      gene targeting and RNA interference.
FAU - Kim, Young June
AU  - Kim YJ
AD  - Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for 
      Regenerative Medicine, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 
      31116, South Korea.
AD  - Institute of Green Bioscience and Technology, Seoul National University, 
      Pyeongchang, 25354, South Korea.
FAU - Kim, Tae-Hyun
AU  - Kim TH
AD  - Institute of Tissue Regeneration Engineering, Dankook University, Cheonan, 31116, 
      South Korea.
FAU - Kim, Minjeong
AU  - Kim M
AD  - Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for 
      Regenerative Medicine, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 
      31116, South Korea.
FAU - Kim, Min Ju
AU  - Kim MJ
AD  - Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for 
      Regenerative Medicine, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 
      31116, South Korea.
FAU - Kim, Hae-Won
AU  - Kim HW
AD  - Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for 
      Regenerative Medicine, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 
      31116, South Korea.
AD  - Institute of Tissue Regeneration Engineering, Dankook University, Cheonan, 31116, 
      South Korea.
FAU - Shim, Hosup
AU  - Shim H
AD  - Department of Nanobiomedical Science and BK21 PLUS NBM Global Research Center for 
      Regenerative Medicine, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 
      31116, South Korea. shim@dku.edu.
AD  - Institute of Tissue Regeneration Engineering, Dankook University, Cheonan, 31116, 
      South Korea. shim@dku.edu.
AD  - Department of Physiology, School of Medicine, Dankook University, Cheonan, 31116, 
      South Korea. shim@dku.edu.
LA  - eng
PT  - Journal Article
DEP - 20160503
PL  - Germany
TA  - In Vitro Cell Dev Biol Anim
JT  - In vitro cellular & developmental biology. Animal
JID - 9418515
RN  - 0 (Tumor Suppressor Protein p53)
SB  - IM
MH  - Animals
MH  - Fibroblasts/metabolism
MH  - Gene Expression Regulation/*genetics
MH  - Gene Knockout Techniques
MH  - Gene Targeting
MH  - Heterozygote
MH  - Humans
MH  - Mice
MH  - Mutation
MH  - *RNA Interference
MH  - Swine
MH  - Tumor Suppressor Protein p53/*biosynthesis/genetics
OTO - NOTNLM
OT  - Knockdown
OT  - Knockout
OT  - Pig
OT  - p53
EDAT- 2016/05/05 06:00
MHDA- 2017/03/03 06:00
CRDT- 2016/05/05 06:00
PHST- 2016/01/05 00:00 [received]
PHST- 2016/04/03 00:00 [accepted]
PHST- 2016/05/05 06:00 [entrez]
PHST- 2016/05/05 06:00 [pubmed]
PHST- 2017/03/03 06:00 [medline]
AID - 10.1007/s11626-016-0026-0 [pii]
AID - 10.1007/s11626-016-0026-0 [doi]
PST - ppublish
SO  - In Vitro Cell Dev Biol Anim. 2016 Aug;52(7):736-41. doi: 
      10.1007/s11626-016-0026-0. Epub 2016 May 3.