scholarly journals Targeting Expression of the Leukemogenic PML-RARα Fusion Protein by Lentiviral Vector-Mediated Small Interfering RNA Results in Leukemic Cell Differentiation and Apoptosis

2011 ◽  
Vol 22 (12) ◽  
pp. 1593-1598 ◽  
Author(s):  
Simone V. Ward ◽  
Thomas Sternsdorf ◽  
Niels-Bjarne Woods
Keyword(s):  
Cell Differentiation ◽  
Fusion Protein ◽  
Small Interfering Rna ◽  
Lentiviral Vector ◽  
Leukemic Cell ◽  
Interfering Rna

scholarly journals Optimization Procedure for Small Interfering RNA Transfection in a 384-Well Format

2007 ◽  
Vol 12 (4) ◽  
pp. 546-559 ◽  
Author(s):  
Jason Borawski ◽  
Alicia Lindeman ◽  
Frank Buxton ◽  
Mark Labow ◽  
L. Alex Gaither
Keyword(s):  
High Throughput Screening ◽  
Mammalian Cells ◽  
Small Interfering Rna ◽  
Dna Analysis ◽  
Lentiviral Vector ◽  
Mrna Levels ◽  
Sirna Transfection ◽  
Rna Transfection ◽  
Well Assay ◽  
Interfering Rna

High-throughput screening of RNAi libraries has become an essential part of functional analysis in academic and industrial settings. The transition of a cell-based RNAi assay into a 384-well format requires several optimization steps to ensure the phenotype being screened is appropriately measured and that the signal-to-background ratio is above a certain quantifiable threshold. Methods currently used to assess small interfering RNA (siRNA) efficacy after transfection, including quantitative PCR or branch DNA analysis, face several technical limitations preventing the accurate measurement of mRNA levels in a 384-well format. To overcome these difficulties, the authors developed an approach using a viral-based transfection system that measures siRNA efficacy in a standardized 384-well assay. This method allows measurement of siRNA activity in a phenotypically neutral manner by quantifying the knockdown of an exogenous luciferase gene delivered by a lentiviral vector. In this assay, the efficacy of a luciferase siRNA is compared to a negative control siRNA across many distinct assay parameters including cell type, cell number, lipid type, lipid volume, time of the assay, and concentration of siRNA. Once the siRNA transfection is optimized as a 384-well luciferase knockdown, the biologically relevant phenotypic analysis can proceed using the best siRNA transfection conditions. This approach provides a key technology for 384-well assay development when direct measurement of mRNA knockdown is not possible. It also allows for direct comparison of siRNA activity across cell lines from almost any mammalian species. Defining optimal conditions for siRNA delivery into mammalian cells will greatly increase the speed and quality of large-scale siRNA screening campaigns. ( Journal of Biomolecular Screening 2007:546-559)

scholarly journals Fusion protein–mediated small interfering RNA delivery

2009 ◽  
Vol 2 (21) ◽  
pp. 892-892
Keyword(s):  
Fusion Protein ◽  
Small Interfering Rna ◽  
Interfering Rna

Random small interfering RNA library screen identifies siRNAs that induce human erythroleukemia cell differentiation

2011 ◽  
Vol 52 (3) ◽  
pp. 502-514 ◽  
Author(s):  
Cuiqing Fan ◽  
Yuan Xiong ◽  
Ning Zhu ◽  
Yabin Lu ◽  
Jiewen Zhang ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Small Interfering Rna ◽  
Library Screen ◽  
Erythroleukemia Cell ◽  
Interfering Rna

Prion or virino or just a small interfering RNA?

2004 ◽  
Author(s):  
Y. E. Karapetyan
Keyword(s):  
Small Interfering Rna ◽  
Interfering Rna
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