Artistic 3D rendition of a circular plasmid

Expand your experimental possibilities beyond synthetic siRNAs using vector-based expression of target-specific shRNA and miRNA. Achieve:

  • Stable or transient target knockdown
  • Lentiviral delivery in hard-to-transfect, primary, and non-dividing cells
  • Co-expression of a fluorescent reporter for visualization

RNAi vector technologies allow you to:

  • Regulate gene inhibition with inducible RNAi expression
  • Select for a pure population of cells stably expressing an RNAi sequence
  • Control gene expression in vivo with tissue-specific promoters


Summary of Invitrogen RNAi vector technologies

 BLOCK-iT Pol II miR RNAi
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BLOCK-iT shRNA
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DescriptionmicroRNA-adapted inserts for more efficient processing of the RNAi moleculeTraditional short hairpin inserts for transient or long-term RNAi
Promoter typePol IIPol III
# shRNA per vectorPolycistronic expression of miR-adapted shRNAs (chaining)Single shRNA expression per promoter
Expression trackingCo-expression of GFP allows expression to be trackedDelivery can be tracked with a separate cassette but not by shRNA expression
Vector compatibilityCompatible with most Gateway destination vectorsCompatible with BLOCK-iT destination vectors
Products

Pre-designed and custom miR RNAi vector inserts
Lentiviral, inducible, and GFP-expressing options available

 

Custom shRNA vectors for any organism

Lentiviral and adenoviral options available

Highly efficient delivery of BLOCK-iT Pol II miR RNAi Expression Vectors or BLOCK-iT shRNA Vectors is necessary to achieve significant levels of knockdown. Optimizing transfection or transduction, controlling for experimental variability, and using a powerful transfection reagent vastly improves the chances for RNAi success. The use of positive and negative controls will also help in the assessment of your experiments.

Optimizing delivery conditions

  • Start with the cell-type specific protocol and transfection reagent most suitable for your experiment
  • Use a DNA plasmid that expresses a fluorescent protein, such as GFP, to monitor that the plasmid has entered the cell
  • Apply a viability indicator such as Ethidium Homodimer-1 (EthD-1) to monitor cell viability related to transfection conditions
  • Utilize Hoechst nuclei stain to measure the percent of transfected and/or dead cells to the total cell population

Resources

Questions

Send your questions to our RNAi team at RNAiSupport@thermofisher.com