SMARTvector Lentiviral shRNA

Make smart, informed choices for successful gene silencing in your cells of interest


Guaranteed gene silencing with state-of-the-art shRNA designs and multiple promoter and reporter options.

Enter a common gene identifier and click SEARCH to find products for your gene.

 
Individual SMARTvector Human Lentiviral shRNA
V3SH7590 100 µL, 10^8 TU/mL - $1,195.00select
Individual SMARTvector Mouse Lentiviral shRNA
V3SM7592 100 µL, 10^8 TU/mL - $1,195.00select
Individual SMARTvector Rat Lentiviral shRNA
V3SR7594 100 µL, 10^8 TU/mL - $1,195.00select
Set of 3 SMARTvector Human Lentiviral shRNA
V3SH7596 100 µL, 10^8 TU/mL - $1,995.00select
Set of 3 SMARTvector Mouse Lentiviral shRNA
V3SM7598 100 µL, 10^8 TU/mL - $1,995.00select
Set of 3 SMARTvector Rat Lentiviral shRNA
V3SR7600 100 µL, 10^8 TU/mL - $1,995.00select
Individual SMARTvector Human LNC shRNA
V3SH7945 100 µL, 10^8 TU/mL - $1,195.00select
Individual SMARTvector Mouse LNC shRNA
V3SM7947 100 µL, 10^8 TU/mL - $1,195.00select
Individual SMARTvector Rat LNC shRNA
V3SR7949 100 µL, 10^8 TU/mL - $1,195.00select
Set of 3 SMARTvector Human LNC shRNA
V3SH7951 100 µL, 10^8 TU/mL - $1,995.00select
Set of 3 SMARTvector Mouse LNC shRNA
V3SM7953 100 µL, 10^8 TU/mL - $1,995.00select
Set of 3 SMARTvector Rat LNC shRNA
V3SR7955 100 µL, 10^8 TU/mL - $1,995.00select

SMARTvector Lentiviral shRNA offers maximum flexibility for vector-based RNAi experiments. Because efficient gene silencing depends on both the design and level of expression of the shRNA, it is critical to choose silencing reagents where both the targeting sequence and the specific promoter driving expression are taken into consideration.

SMARTvector Lentiviral shRNAs are designed using our most advanced microRNA-based shRNA-specific rational design algorithm resulting in highly potent and specific targeting sequences. Choose from seven promoters (human and mouse CMV, human and mouse EF1 alpha, CAG, PGK, and UBC), two fluorescent reporters (TurboGFP and TurboRFP) and a no-fluorescent reporter option to tailor SMARTvector lentiviral shRNAs for your specific cells of interest and your experimental requirements. Up to ten shRNA silencing constructs are predesigned for every gene in human, mouse and rat providing broad coverage across the entire genome.

Unsure which format you need? Click here to learn about our SMARTchoice configuration platform.

Highlights

SMARTvector Lentiviral shRNA

  • Target any gene in human, mouse and rat, and tailor experiments to specific cells with multiple promoter options
  • Guaranteed silencing (see Specifications tab)
  • Designed using microRNA scaffold-specific attributes for highly efficient processing via the endogenous RNAi pathway
  • Available as glycerol stock and as purified, concentrated lentiviral particles
  • Standard titers of 1 x 108 TU/mL, ± 20%, and ultra-high titers of 5 x 109 TU/mL, ± 20%; functional titers determined by flow cytometric analysis of GFP-positive, transduced HEK293T cells
  • Suitable for dividing and non-dividing cell types, including difficult-to-transfect cells such as primary, neuronal and stem cells

Together, these attributes greatly enhance the functionality and specificity of lentiviral-mediated RNAi and reduce the toxicity associated with low titer preparations (supernatant).

Superior Dharmacon design algorithm

  • Utilization of a highly processed, patented microRNA scaffold
  • Rationally-designed, highly functional gene targeting sequences
  • Algorithm trained on more than 500 endogenous mRNA data points for functional knockdown testing
  • Incorporation of bioinformatics strategies to reduce off-target gene knockdown events
  • Protein coding genes are targeted in the ORF and 3’UTR; lncRNAs are targeted throughout the entire transcript length

Lentiviral particle formats

At Dharmacon, we are committed to delivering high quality lentiviral particle preparations. As such, we clone, package and concentrate each construct with strict QC controls. Due to the inherent complexity of lentiviral particle production and the quality control procedures, turnaround time is estimated to be 4 to 6 weeks. We offer the following formats:

Set of 3 different gene-targeting constructs

  • Ideal for evaluating multiple shRNAs targeting the same gene and ensuring mRNA knockdown in your specific cells
  • 100 µL (4 x 25 µL) or 200 µL (8 x 25 µL) total amounts at 1 x 108 TU/mL, ± 20%, functional titers determined by flow cytometric analysis of GFP-positive, transduced HEK293T cells

Individual gene-targeting constructs

  • Ideal for ordering one or two constructs that you have identified as being the best performers
  • 100 µL (4 x 25 µL) or 200 µL (8 x 25 µL) total amounts at 1 x 108 TU/mL, ± 20%, or 100 µL (4 x 25 µL) total amount at 5 x 109 TU/mL, ± 20%, functional titers determined by flow cytometric analysis of GFP-positive, transduced HEK293T cells

SMARTvector Lentiviral Positive and Negative Controls for shRNA and microRNA experiments

  • Ideal for optimizing transduction and assay conditions prior to gene-specific experimentation
  • Negative control particles allow for distinguishing sequence-specific silencing from non-specific effects
  • Positive control particles allow for confirmation of gene silencing using confirmed shRNA constructs targeting a housekeeping gene
  • 50 µL (2 x 25 µL) at 1 x 108 TU/mL, ± 20% or 100 µL (4 x 25 µL) at 5 x 109 TU/mL, ± 20%, functional titers determined by flow cytometric analysis of GFP-positive, transduced HEK293T cells
  • Click here to learn about SMARTvector controls
  
HazardousNo
Shipping ConditionDry Ice,Wet Ice
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition-80 C

SMARTvector Lentiviral shRNA functional guarantee

When you purchase a minimum of three SMARTvector lentiviral shRNAs to the same protein-coding gene target using the optimal SMARTchoice promoter for your cell type, at least one of the shRNA constructs will reduce target mRNA levels by 70% or more when used with the vector matched non-targeting control and GAPD or PPIB positive control. Optimal promoters should be determined with the SMARTchoice promoter selection plate. mRNA levels should be measured with RT-qPCR or similar quantitative mRNA analysis no earlier than 72 hours post-transduction. The guarantee only applies to catalog numbers V3S*xxxx and does not apply to any SMARTvector shRNAs targeting lncRNA transcripts.

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SMARTvector Lentiviral shRNA vector elements

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Vector Element Utility
5' LTR 5' Long Terminal Repeat necessary for lentiviral particle production and integration of the construct into the host cell genome
Ψ Psi packaging sequence allows viral genome packaging using lentiviral packaging systems
PRRE Rev Response Element enhances titer by increasing packaging efficiency of full-length viral genomes
tGFP or tRFP TurboGFP or TurboRFP reporter for visual tracking expression upon doxycycline induction
None No-reporter option for use in applications where fluorescence is not required or desired
IRES Internal Ribosomal Entry Site allows expression of fluorescent marker and puromycin resistance in a single transcript
PuroR Puromycin resistance permits antibioticselective pressure and propagation of stable integrants
SMARTvector universal scaffold Optimized proprietary scaffold in which mature microRNA sequence is embedded
WPRE Woodchuck Hepatitis Post-transcriptional Regulatory Element enhances transgene expression in target cells
3' SIN LTR 3' Self-inactivating Long Terminal Repeat for increased Lentiviral safety


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Functionality of endogenous microRNAs are different

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Incorporation of highly functional and appropriately processed microRNA scaffolds into lentiviral silencing constructs greatly enhances the functionality of DNA-based RNAi silencing. Different endogenous human microRNAs were tested in HeLa cells for the ability to silence their respective targets. Each microRNA exhibits a distinctive level of functionality. Several of the constructs (e.g., miR-486 and -526a) exhibited poor levels of activity, silencing their respective reporter constructs less than 20% while native miR-30 and -374 scaffolds provide moderate levels of gene silencing (~50-70%, see above). The scaffold selected as the basis for SMARTvector Lentiviral shRNAs efficiently processed and silenced its’ respective target by 80 to 90%. This highly functional microRNA represents a significant improvement over currently available microRNA-based scaffolds used in RNAi expression platforms.


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Functionality of mature and passenger strands of endogenous microRNAs are different

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Preferred microRNA scaffolds have minimal passenger strand off-target effects. The Dharmacon team investigated the passenger strand activity of several of the scaffolds tested in the primary screen. The moderately functional miR-30 and the highly functional miR-126 exhibit undesirable passenger strand activity. In contrast, miR-SMARTvector shows potent mature strand activity with minimal passenger strand function.


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microRNA-adapted shRNAs compared to simple hairpin shRNAs

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The SMARTvector microRNA-adapted shRNA platform shows considerably higher mature strand activity and lower passenger stand activity than simple hairpin shRNAs targeting the same gene. The result of the optimized microRNA-adapted scaffold is higher, more specific function while reducing the risk of off-target effects from the passenger strand.


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SMARTvector lentiviral shRNA particles in two cell lines

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Two different cell lines were transduced with Dharmacon SMARTvector Lentiviral shRNA Particles targeting CDK9, HIF1α, ILK, MAPK1 or PTEN (as indicated) at three MOIs. mRNA knockdown was assessed 72 hours post-transduction using QuantiGene branched DNA assay (Panomics, Inc.) and normalized to SMARTvector Non-targeting Control. In most cases, 70% or greater gene knockdown was achieved in the absence of puromycin selection. Rational design, broad tropism, and GFP reporter expression facilitate transduction optimization that leads to efficient gene knockdown in cells refractory to lipid-mediated delivery.


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The SMARTvector no reporter option provides equivalent gene knockdown compared to the SMARTvector constructs with a fluorescent reporter

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A549 cell lines were transduced with Dharmacon SMARTvector Lentiviral shRNA Particles targeting GAPD with and without a fluorescent reporter at a range of MOIs. mRNA knockdown was assessed 5 days after puromycin selection using QuantiGene branched DNA assay (Panomics, Inc.) and normalized to the matched SMARTvector Non-targeting Control. In all cases, GAPD knockdown is the similar with and without fluorescent reporter.