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

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 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

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.

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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

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

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

Products for Step 5: Correlate