TransIT-siQUEST® Transfection Reagent

A high efficiency, low toxicity, siRNA transfection reagent for mammalian cells

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MIR 21140.4 mlMIR 21101.5 mlMIR 21155 x 1.5 mlMIR 211610 x 1.5 ml

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  • Broad Spectrum siRNA Delivery – Utilize one transfection reagent and protocol for a wide variety of cells
  • Low Cellular Toxicity – Maintain cell density and reduce experimental biases
  • Reproducible Results – Obtain consistent, targeted gene knockdowns from day to day
  • High Knockdown Efficiency – Achieve optimal gene silencing in a large percentage of cells to ensure experimental success

Figures and Data

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High Knockdown and Low Toxicity Using TransIT-siQUEST Reagent in CHO Cells Stably Expressing Firefly Luciferase

High Knockdown and Low Toxicity Using TransIT-siQUEST® Reagent in CHO Cells Stably Expressing Firefly Luciferase. CHO-luc cells were grown in 24 wells plates and transfected with 25 nM of either a non-targeting siRNA or a anti-firefly luciferase siRNA using the indicated reagents with the volumes noted beneath each well. Luciferase expression, normalized to non-targeting siRNA control (bar graph) and lactate dehydrogenase (LDH) levels (line graph) were measured at 24 hours post-transfection. LDH levels are reported as percent cytotoxicity compared to cells alone and were measured using a commercially available colorimetric assay; all values at or below zero are represented as zero on graph.

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Inhibition of PPAR-alpha Expression in Primary Mouse Hepatocytes Using the TransIT-siQUEST Reagent

Inhibition of PPAR-alpha Expression in Primary Mouse Hepatocytes Using the TransIT-siQUEST® Reagent. Primary mouse hepatocytes were transfected with an anti-PPAR-alpha siRNA or a non-targeting control siRNA using the TransIT-siQUEST® Reagent. Twenty-four hours post-transfection, the amount of PPAR-alpha mRNA was measured relative to GAPDH mRNA levels using qRT-PCR and then scaled to the expression level of the specific target mRNA in the Opti-MEM® media only (untreated) control.

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Delivery of Fluorescently-Labeled siRNA Using TransIT-siQUEST Transfection Reagent

Delivery of Fluorescently-Labeled siRNA Using TransIT-siQUEST® Transfection Reagent. HeLa (70% confluence) cells in 12-well plates were transfected with TransIT-siQUEST® Transfection Reagent (3 μl/well) and Label IT® siRNA Tracker™ Cy-3-labeled siRNA duplexes (RED, 25 nM final concentration in the well). The cells were incubated 24 hours post-transfection then fixed and counterstained with counterstained with Alexa Fluor® 488 Phalloidin (GREEN) (Life Technologies). Confocal images were acquired on a Zeiss LSM 510 Confocal Microscope.

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Efficient Target Gene Knockdown in Selected Cell Lines Using TransIT-siQUEST Reagent

Efficient Target Gene Knockdown in Selected Cell Lines Using TransIT-siQUEST® Reagent. Reporter plasmids expressing both firefly and sea pansy luciferase were co-transfected into the indicated cell lines using TransIT® Plasmid Transfection Reagents. Targeted knockdown was achieved by transfection of an anti-firefly luciferase siRNA using the TransIT-siQUEST® Reagent. Twenty-four hours later, firefly luciferase expression was normalized to sea pansy luciferase expression and compared to the reagent alone control.



Storage Conditions:
All Configurations: Store at 4°C

Product Guarantee:
All Configurations: 1 year

Usage Statement:
All Configurations: For Research Use Only.

Animal Origin Statement:
All Configurations: This product is animal origin free.

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In our hands, we achieved greater than 80% target knockdown and observed lower toxicity when transfecting NCI-295R cells with TransIT-siQUEST® than with Lipofectamine®. Results we obtained using TransIT-siQUEST® are included in our recent publication demonstrating that the nuclear antigen-associated factor KIAA0101 is implicated in adrenal cancer growth and invasion. (Jain, M et al. PLoS ONE. 2011;6(11):e26866. Epub 2011 Nov 11)
Dr. Meenu Jain
Laboratory of Electron Kebebew - National Institutes of Health
Our lab effectively carried out an RNAi screen against 45 mitochondrial candidate genes with TransIT-siQUEST® Transfection Reagent in HEK 293T cells. This work enabled us to identify a critical regulator (MCUR1) for calcium uptake in the mitochondria that regulates cellular metabolism. (Mallilankaraman, K et al. Nature Cell Biology. December 2012)
Dr. Karthik Mallilankaraman
Center for Translational Medicine - Temple University (Madesh Lab)
Our lab routinely uses Mirus Bio transfection products, and we recently published using TransIT-siQUEST® to perform RNA interference experiments in LNCaP cells to elucidate the role of p53 in selenium-induced apoptosis in prostate cancer cells. These results were published in 2010 in the International Journal of Oncology. (Sarveswaran, S. et al. IJO. Jan 2010)
Dr. Jagadananda Ghosh
Henry Ford Health System - Vattikuti Urology Institute