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Author: Wu, H
Author: Mo, Y
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Methods Article

Generation of shRNAs from randomized oligonucleotides

Hailong Wu1, Anh Dinh1 and Yin-Yuan Mo1*

1 Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine. Springfield, IL 62794. USA.

* To whom correspondence should be addressed: Yin-Yuan Mo, Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University. 801 N. Rutledge PO Box 19626, Springfield, IL 62794. USA. Phone: 217-545-8505. Fax: 217-545-7569. Email: ymo@siumed.edu

Biol. Proced. Online 2007;9:9-17. doi:10.1251/bpo129
Submitted: February 15, 2006; Accepted: March 28, 2007; Published: July 04, 2007.

Indexing terms: RNA Interference; RNA, Small Interfering; Oligonucleotides.

Abbreviations: bp, base pair; RNAi, RNA interference; shRNA, short hairpin RNA; siRNA, short interfering RNA.


Figure 3 Enlarged

Fig. 3:

Suppression of p53 by p53-shRNA. A: Detection of p53 expression by Western blot. Plasmid carrying an appropriate DNA fragment was introduced into MCF-7 cells. Proteins samples were prepared as described in Materials and Methods. Lanes 1, vector control; 2, p53-shRNA/pSK-H1-Pu-X; 3, p53-shRNA-p (positive control) and 4, p53-shRNA/pSK-H1-Pu-X/Xho I. β-actin serves as a loading control. B: Removal of siRNA-loop-1 in p53-shRNA. Both constructs were digested with BamH I and Kpn I. M, 25 bp DNA ladders; lanes 1, p53-shRNA/pSK-H1-Pu-X; 2, p53-shRNA/pSK-H1-Pu-X/Xho I. C: Sequence comparison of p53-shRNA/pSK-H1-Pu-X and p53-shRNA/pSK-H1-Pu-X/Xho I. The p53 specific sequence is underlined.

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