A novel RNAi library based on partially randomized consensus sequences of nuclear receptors: identifying the receptors involved in amyloid beta degradation

Genomics
Jian-Ping YangQi-Xiang Li

Abstract

Combinatorial gene inactivation using an RNAi library is a powerful approach to discovering novel functional genes. However, generation of a comprehensive RNAi library remains technically challenging. In this report, we describe a simple and novel approach to designing gene-family-specific RNAi libraries by targeting conserved motifs using degenerate oligonucleotides. We created an siRNA library in the pHUMU vector using partially randomized sequences targeting the consensus region in the ZnF_C4 signature motif of the nuclear hormone receptors and thus against the entire receptor superfamily. For proof of principle, we adapted a reporter assay to screen this library for receptors that might be involved in reducing amyloid beta peptide accumulation. We modified a previously described luciferase reporter assay to measure the amyloid beta precursor cleavages occurring only between beta- and gamma-secretase cleavage sites, thus excluding the major gamma-secretase activities that could generate neurotoxic Abeta peptides. Our screen using this assay identified siRNA vectors that specifically increase the Abeta40/42 cleavage and pointed to a potential receptor target, ROR-gamma. SiRNAs targeting other regions of ROR-gamma not only con...Continue Reading

References

Dec 30, 1994·Biochemical and Biophysical Research Communications·T HiroseA M Jetten
Jul 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·M KrugerF Wong-Staal
Jan 3, 2001·Proceedings of the National Academy of Sciences of the United States of America·C BegerF Wong-Staal
Apr 17, 2003·Proceedings of the National Academy of Sciences of the United States of America·Bryan A LaffittePeter Tontonoz
Jun 26, 2003·The International Journal of Biochemistry & Cell Biology·Yan LingNoor Kalsheker
Oct 22, 2003·Cell·Dianne S SchwarzPhillip D Zamore
Oct 23, 2003·Nature Reviews. Cancer·Thijn R Brummelkamp, René Bernards
Dec 23, 2003·Proceedings of the National Academy of Sciences of the United States of America·Lianxing ZhengPeter G Schultz
Jan 6, 2004·Nature Genetics·George SenHelen M Blau
Jan 6, 2004·Nature Genetics·Daisuke ShiraneKenzo Hirose
Feb 18, 2004·Current Pharmaceutical Biotechnology·J Zhang, Z C Hua
Mar 17, 2004·Proceedings of the National Academy of Sciences of the United States of America·Biao LuoHarvey F Lodish
Mar 26, 2004·Nature·Patrick J PaddisonGregory J Hannon
Nov 3, 2004·Nature Reviews. Drug Discovery·Hinrich GronemeyerVincent Laudet
Mar 7, 2006·Biochemical and Biophysical Research Communications·Guohong LiuQi-Xiang Li

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Citations

Sep 20, 2012·BMC Genomics·Chih-Hung ChangChuan Yi Tang
Jan 19, 2008·Cell Death and Differentiation·N M Wolters, J P MacKeigan
Oct 1, 2006·Drug Discovery Today. Technologies·Guohong LiuQi-Xiang Li
Aug 28, 2007·Neurochemistry International·Neville Marks, Martin J Berg
Sep 16, 2020·BMC Genomics·Natalia RubanovaNadya Morozova

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