Brief Article
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World J Gastroenterol. Oct 21, 2010; 16(39): 4980-4985
Published online Oct 21, 2010. doi: 10.3748/wjg.v16.i39.4980
CABYR RNAi plasmid construction and NF-κB signal transduction pathway
Lin-Xiang Shi, Yao-Ming He, Lin Fang, Hong-Bo Meng, Li-Jun Zheng
Lin-Xiang Shi, Yao-Ming He, Lin Fang, Hong-Bo Meng, Li-Jun Zheng, Department of General Surgery, Shanghai Tenth People’s Hospital, Tongji University, Shanghai 200072, China
Author contributions: Shi LX and He YM performed the majority of experiments; Fang L provided the vital reagents and financial support for this work; Fang L and He YM designed the study; He YM, Meng HB and Zheng LJ were involved in editing the manuscript and analytical tools.
Supported by Grants from Natural Science Foundation of China, No. 30940034
Correspondence to: Lin Fang, MD, PhD, Department of General Surgery, Shanghai Tenth People’s Hospital, Tongji University, Shanghai 200072, China. ajijinwei@126.com
Telephone: +86-21-66301059 Fax: +86-21-66301051
Received: May 1, 2010
Revised: June 10, 2010
Accepted: June 17, 2010
Published online: October 21, 2010
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.

Keywords: CABYR, Plasmid, Nuclear factor-κB, Signal transduction, RNAi, Cabyrmid 2