Gene Knockdown | Life Technologies

Gene Knockdown

Post-translational silencing of gene expression is a powerful molecular tool for stem cell research to study gene function, biological pathways and physiology of disease. Gene silencing via siRNA enables several applications:

Developmental pathway screening to determine the gene function and identify transcription factors that regulate self-renewal, proliferation, differentiation and apoptosis.
Directed differentiation of embryonic or tissue-specific stem cells by silencing genes of pluripotency maintenance, such as Oct 4 or Nanog to induce early differentiation and lineage determination.
Reprogram or dedifferentiate tissue specific stem cells to repair damaged tissues or organs.

The Life Technologies siRNA Analysis Workflow is an optimized system for inducing gene silencing. Benefits include:

Integrated workflow: Solutions for siRNA selection, transfection, validation and functional analysis by real-time PCR
GeneAssist™ Pathway Atlas: An interactive on-line tool to select siRNAs and assays within the context of biological process
Silencer® Select Pre-designed siRNAs: Chemically modified siRNAs with improved silencing efficacy, potency and specificity that minimize artifacts due to poor silencing and off-target effects

News

Workflow
Publications & Literature

Experimental Workflow: Find the Products You Need for Every Step

Using the right siRNAs is important for gene knockdown and Applied Biosystems is pleased to offer Silencer® Select Pre-Designed siRNAs or Custom Designed Silencer® Select siRNAs, chemically modified siRNAs with improved silencing efficacy, potency and specificity that minimize artifacts due to poor silencing and off-target effects.

Products for Step 1: Select siRNAs

Silencer® Select Pre-Designed and Validated siRNA incorporate the latest improvements in siRNA design, off-target effect prediction algorithms, and chemical modifications to yield siRNA with unrivalled efficacy, potency, and specificity. The result is fewer failed experiments due to poor silencing, and cleaner, more consistent phenotypic data.

This product is not orderable online.

For more information, please contact your local sales representative.

For siRNA transfection, Applied Biosystems recommends Invitrogen Lipofectamine RNAiMAX, a proprietary RNAi-specific cationic lipid formulation that offers the highest transfection efficiencies on the widest variety of cell types for siRNA gene knockdown experiments.

Products for Step 2: Deliver

Lipofectamine RNAiMAX

learn more

Single-tube lysis and RT is performed using the TaqMan® Gene Expression Cells-to-CT™ Kit. For single cell analysis, TaqMan® PreAmp Cells-to-CT™ Kit is recommended. TaqMan® Stem Cell Pluripotency Arrays are used to characterize undifferentiated stem cells or their derivatives. To characterize embryonic or lineage-specific stem cells or assess gene targets in specific pathways, TaqMan® Gene Expression Assays can be used. The TaqMan® Gene Expression Assays are compatible with Applied Biosystems Real-Time PCR Instruments listed below.

Products for Step 3: Test Efficiency

TaqMan® Gene Expression Cells-to-CT™ Kit

This kit enables researchers to easily perform expression analysis using cultured cells without purification of RNA. The kit includes reagents for cell lysis and RNA preservation, genomic DNA removal, and reverse transcription (RT). The kit also includes TaqMan® Gene Expression Master Mix for use in subsequent real-time PCR analysis.

TaqMan® PreAmp Cells-to-CT™ Kit

Perform gene expression analysis directly from limited or small numbers of cultured cells without RNA purification. The protocol includes an intermediate amplification step (pre-amplification) between reverse transcription and real-time PCR in which cDNA is enriched for up to 100 gene targets of the user’s choice without amplification bias.

TaqMan® Stem Cell Pluripotency Array

The Human Stem Cell Pluripotency Array contains a well-defined set of validated gene expression markers to characterize embryonic stem (ES) cell identity and assess phenotypic variations between embryonic stem cell isolates.

TaqMan® Gene Expression Assays

Life Technologies offers more than 1,000,000 TaqMan® Gene Expression Assays, the most comprehensive set of pre-designed Real-Time PCR assays available. All TaqMan Gene Expression Assays have been designed using our validated bioinformatics pipeline, and run with the same PCR protocol, eliminating the need for primer design or PCR optimization.

search for assays

TaqMan® Gene Expression Master Mix

The TaqMan® Gene Expression Master Mix is tailored for precise quantification by real-time PCR. It is designed for routine and challenging experiments.

StepOnePlus™ Real-Time PCR System

The Applied Biosystems® StepOnePlus™ Real-Time PCR System makes it simple and easy to get publication-ready high-quality real-time PCR results on day one. This remarkably simple 96-well, 4-color, real-time PCR system is designed with a user-friendly, yet powerful, interface for researchers of all experience levels.

7500 Real-Time PCR System

The Applied Biosystems® 7500 Real-Time PCR System is a powerful platform for labs requiring superior performance and maximum dye versatility. This third-generation platform features an innovative optical system that enhances sensitivity and lets you access a broader range of fluorophores. The variable excitation capability allows greater sensitivity for longer wavelength (red) dyes.

7500 Fast Real-Time PCR System

The Applied Biosystems® 7500 Fast Real-Time PCR System offers maximum performance in the minimum time. Fully optimized for Fast cycling, the 7500 Fast System delivers high-quality results in as little as 30 minutes.

7900HT Fast Real-Time PCR System

The 7900HT Fast Real-Time PCR System is the only real-time quantitative PCR system that combines 384-well plate compatibility with fully automated robotic loading — and also offers optional Fast real-time PCR capability. Acknowledged as the gold standard in real-time PCR, the 7900HT System combined with TaqMan® Genomic Assays enables you to achieve unprecedented throughput and flexibility, allowing you to pursue projects beyond the scope of previous real-time instruments.

View ordering information

Assess the phenotypic effects by examining cell morphology, measuring mRNA with TaqMan® Gene Expression Workflow or protein levels with the Western-Star™ Immunodetection System.

Products for Step 4: Examine Biologically

TaqMan® Protein Assays

TaqMan® Protein Assays help quantitate protein expression using gold standard TaqMan® 5' nuclease chemistry with a workflow and sample quantity similar to our TaqMan® Gene Expression and MicroRNA (miRNA) Assays. As the protein expression results are obtained on the same analytical platform, these revolutionary assays enable direct correlation of mRNA and/or miRNA expression to protein expression.

Western-Star™ Immunodetection System

Western-Star™ system is a versatile, non-isotopic chemiluminescent immunodetection system.

View ordering information

Compare phenotype and level of gene knockdown using time course or siRNA titration studies.

Products for Step 5: Correlate

TaqMan® Protein Assays

*Note: Running a complete TaqMan® Protein Assay (pre-designed or made with the Open Kit) requires a Core Reagents Base Kit and Master Mix Kit.

View ordering information

References

Guides