CABYR RNAi plasmid construction and NF-κB signal transduction pathway

World J Gastroenterol. 2010 Oct 21;16(39):4980-5. doi: 10.3748/wjg.v16.i39.4980.

Abstract

Aim: To construct the CABYR RNAi plasmid and study its relation with the nuclear factor (NF)-κB signal transduction pathway.

Methods: Human CABYR mRNA sequence was obtained from GenBank. The structure of cDNA sequence for the short hairpin RNA was BbsI + sense + loop + antisense + transcription terminator + KpnI + BamHI. A CABYR silencing plasmid was constructed and transfected into the human embryo cell line 293T. Quantitative real-time polymerase chain reaction was used to analyze CABYR and NF-κB gene expression.

Results: The CABYR and NF-κB expressions were detected in 293T cells. The oligonucleotide (5'-GCTCAGATGTTAGGTAAAG-3') efficiently silenced the expression of CABYR. The expression of NF-κB was not significantly affected by silencing CABYR (P = 0.743).

Conclusion: CABYR can be found in the human embryo cell line 293T. Cabyrmid 2 can efficiently silence its target, CABYR, indicating that CABYR is not related with the NF-κB signal transduction pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism*
  • Cell Line
  • Humans
  • I-kappa B Proteins / metabolism
  • Molecular Sequence Data
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Phosphorylation
  • Plasmids
  • RNA Interference*
  • RNA, Messenger / metabolism
  • Signal Transduction*
  • Transfection

Substances

  • CABYR protein, human
  • Calcium-Binding Proteins
  • I-kappa B Proteins
  • NF-kappa B
  • NFKBIA protein, human
  • Phosphoproteins
  • RNA, Messenger
  • NF-KappaB Inhibitor alpha