shRNA expression constructs designed directly from siRNA oligonucleotide sequences.

Molecular Biotechnology
Tuva BarøyEirik Frengen

Abstract

Gene silencing by RNA interference (RNAi) is a widely used approach for target-specific knockdown of gene expression. Induction of RNAi in mammalian cells can be achieved by introduction of synthetic small interfering RNA (siRNA) resulting in transient knockdown, or alternatively by stable expression of short hairpin RNA (shRNA). Several algorithms for efficient siRNA design exist, but recent reports have suggested that these cannot be directly used to design efficient shRNAs. In this study, 25 siRNAs targeting independent sequences in five transcripts were used for the construction of shRNA cassettes. Both the siRNAs and shRNA constructs were transfected into HEK293T cells. Quantitative real-time PCR analysis revealed that 19 out of the 25 shRNA constructs reduced the average expression level to less than 30%. Our data support that sequences designed by siRNA algorithms efficiently reduce the expression of the target gene when converted into shRNA expression constructs.

References

Oct 31, 2002·Proceedings of the National Academy of Sciences of the United States of America·Luis Alfonso MartinezAnnick Harel-Bellan
Feb 26, 2003·Genes & Development·John G DoenchPhillip A Sharp
Sep 26, 2003·Nature·Amy A CaudyRonald H A Plasterk
Sep 26, 2003·Nature·Yoontae LeeV Narry Kim
Oct 22, 2003·Cell·Anastasia KhvorovaSumedha D Jayasena
Feb 6, 2004·Nature Biotechnology·Angela ReynoldsAnastasia Khvorova
Mar 27, 2004·Biochemical and Biophysical Research Communications·Mohammed Amarzguioui, Hans Prydz
Sep 17, 2004·Nature·Craig C Mello, Darryl Conte
Sep 17, 2004·Nature·Gunter Meister, Thomas Tuschl
Mar 25, 2005·The Journal of Biological Chemistry·Priscilla BragliaGiorgio Dieci
Jul 19, 2005·Nature Biotechnology·Dieter HueskenJonathan Hall
Jan 26, 2006·BMC Biotechnology·Debra J TaxmanWilliam Reed
Aug 25, 2006·Nature Methods·Frank BuchholzMirko Theis
Aug 25, 2006·Nature Methods·René BernardsRoderick L Beijersbergen
Jul 20, 2007·Oligonucleotides·Ute KruegerJuliane Konrad
Jul 27, 2007·Molecular Biotechnology·Gro Leite StørvoldEirik Frengen
Sep 15, 2007·Nature Protocols·Amanda BirminghamAnastasia Khvorova
Mar 29, 2008·Methods in Molecular Biology·Mohammed Amarzguioui, John J Rossi

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