PMID- 26585152
OWN - NLM
STAT- MEDLINE
DCOM- 20160829
LR  - 20230612
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1366
DP  - 2016
TI  - Estradiol-Induced Transcriptional Regulation of Long Non-Coding RNA, HOTAIR.
PG  - 395-412
LID - 10.1007/978-1-4939-3127-9_31 [doi]
AB  - HOTAIR (HOX antisense intergenic RNA) is a 2.2 kb long non-coding RNA (lncRNA), 
      transcribed from the antisense strand of homeobox C (HOXC) gene locus in 
      chromosome 12. HOTAIR acts as a scaffolding lncRNA. It interacts and guides 
      various chromatin-modifying complexes such as PRC2 (polycomb-repressive complex 
      2) and LSD1 (lysine-specific demethylase 1) to the target gene promoters leading 
      to their gene silencing. Various studies have demonstrated that HOTAIR 
      overexpression is associated with breast cancer. Recent studies from our 
      laboratory demonstrate that HOTAIR is required for viability of breast cancer 
      cells and is transcriptionally regulated by estradiol (E2) in vitro and in vivo. 
      This chapter describes protocols for analysis of the HOTAIR promoter, cloning, 
      transfection and dual luciferase assays, knockdown of protein synthesis by 
      antisense oligonucleotides, and chromatin immunoprecipitation (ChIP) assay. These 
      protocols are useful for studying the estrogen-mediated transcriptional 
      regulation of lncRNA HOTAIR, as well as other protein coding genes and non-coding 
      RNAs.
FAU - Bhan, Arunoday
AU  - Bhan A
AD  - Department of Chemistry and Biochemistry, The University of Texas at Arlington, 
      Arlington, TX, 76019, USA.
FAU - Mandal, Subhrangsu S
AU  - Mandal SS
AD  - Department of Chemistry and Biochemistry, The University of Texas at Arlington, 
      Arlington, TX, 76019, USA. smandal@uta.edu.
LA  - eng
GR  - R15 ES019129/ES/NIEHS NIH HHS/United States
GR  - 1R15 ES019129-01/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Estrogens)
RN  - 0 (HOTAIR long untranslated RNA, human)
RN  - 0 (Oligonucleotides, Antisense)
RN  - 0 (RNA, Long Noncoding)
RN  - 4TI98Z838E (Estradiol)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - Breast Neoplasms/genetics/*metabolism
MH  - Chromatin Immunoprecipitation
MH  - Cloning, Molecular
MH  - Dose-Response Relationship, Drug
MH  - Estradiol/*pharmacology
MH  - Estrogens/*pharmacology
MH  - Female
MH  - Gene Expression Regulation, Neoplastic/*drug effects
MH  - Genes, Reporter
MH  - Humans
MH  - Luciferases/genetics/metabolism
MH  - MCF-7 Cells
MH  - Oligonucleotides, Antisense/genetics/metabolism
MH  - Promoter Regions, Genetic
MH  - RNA, Long Noncoding/genetics/*metabolism
MH  - Transcription, Genetic/*drug effects
MH  - Transcriptional Activation/drug effects
MH  - Transfection
MH  - Workflow
OTO - NOTNLM
OT  - Epigenetics
OT  - Estradiol
OT  - Estrogen receptor s
OT  - Gene expression
OT  - HOTAIR
OT  - Long non-coding RNA
OT  - Mixed lineage leukemia
OT  - Polycomb-repressive complex 2
EDAT- 2015/11/21 06:00
MHDA- 2016/08/30 06:00
CRDT- 2015/11/21 06:00
PHST- 2015/11/21 06:00 [entrez]
PHST- 2015/11/21 06:00 [pubmed]
PHST- 2016/08/30 06:00 [medline]
AID - 10.1007/978-1-4939-3127-9_31 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2016;1366:395-412. doi: 10.1007/978-1-4939-3127-9_31.