Specific suppression of beta-secretase gene expression by short interfering RNA in SK-N-SH cells

Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 Nov;37(6):821-4.

Abstract

Objective: To test the effect of short interfering RNAs (siRNAs) of beta-site APP cleaving enzyme (BACE) on inhibiting the expression of BACE in mammalian cells.

Methods: The gene of EGFP, U6 promoter and beta-secretase targeting siRNA were cloned by PCR. The PCR products were inserted into the retrovirus plasmid pLXSN. The interfering vector was identified as pLXSN/ EGFP-U6-siBACE. The SK-N-SH cell line was produced, which can highly expressed BACE. The inhibitive effect of BACE siRNA on BACE expression was examined by fluoroscopy and immunohistochemistry tests.

Results: The interfering vector, pLXSN/EGFP-U6-siBACE, was constructed successfully. The BACE siRNA inhibited the expression of BACE in the SK-N-SH cell and reduced the production of Abeta.

Conclusion: BACE siRNA inhibits the expression of BACE gene of mammalian, which has implications for RNA interference of Alzheimer's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / genetics
  • Amyloid Precursor Protein Secretases / deficiency*
  • Amyloid Precursor Protein Secretases / genetics*
  • Animals
  • Cell Line, Tumor
  • DNA, Recombinant / genetics
  • Gene Expression
  • Genetic Engineering / methods*
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins / genetics
  • Humans
  • Mice
  • Molecular Sequence Data
  • RNA Interference
  • RNA, Small Interfering / genetics*
  • Retroviridae / genetics
  • Retroviridae / physiology
  • Viral Load

Substances

  • DNA, Recombinant
  • RNA, Small Interfering
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Amyloid Precursor Protein Secretases

Associated data

  • GENBANK/U76561
  • GENBANK/X06980
  • RefSeq/NM_012104