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


CONCLUSIONS
BACE siRNA inhibits the expression of BACE gene of mammalian, which has implications for RNA interference of Alzheimer's disease.
Dong
Wei-Jiang
WJ
Department of Anatomy and Histology and Embryology, Xi'an Jiaotong University Medical School, Xi'an 710061, China.
Feng
Gai-Feng
GF
Gong
Hui-Lin
HL
Liu
Su-Hu
SH
Hu
Hai-Tao
HT
D000544:Alzheimer Disease / Q000235:genetics; D053829:Amyloid Precursor Protein Secretases / Q000172:deficiency* / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics / Q000502:physiology; D019562:Viral Load
D004274:DNA, Recombinant; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D053829:Amyloid Precursor Protein Secretases