CABYR RNAi plasmid construction and NF-κB signal transduction pathway.
OBJECTIVE
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).


CONCLUSIONS
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.
Shi
Lin-Xiang
LX
Department of General Surgery, Shanghai Tenth People's Hospital, Tongji University, Shanghai 200072, China. ajijinwei@126.com
He
Yao-Ming
YM
Fang
Lin
L
Meng
Hong-Bo
HB
Zheng
Li-Jun
LJ
D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection
C448922:CABYR protein, human; D002135:Calcium-Binding Proteins; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D010750:Phosphoproteins; D012333:RNA, Messenger; D000072000:NF-KappaB Inhibitor alpha