SMARTvector Lentiviral shRNA
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? Learn about our SMARTchoice configuration platform *Please note that some promoter options may only be available as custom products or upon request.
- Target any gene in human, mouse and rat, and tailor experiments to specific cells with multiple promoter options
- Guaranteed silencing (see Guarantee 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 2 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 2 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 2 x 109 TU/mL, ± 20%, functional titers determined by flow cytometric analysis of GFP-positive, transduced HEK293T cells
- Learn about SMARTvector controls.
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.
- Positive and negative shRNA controls with choice of four promoters and two fluorescent reporters.
- Available for human, mouse and rat.
SMARTvector Lentiviral shRNA vector elements
|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 of expression|
|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|
Functionality of endogenous microRNAs are different
Functionality of mature and passenger strands of endogenous microRNAs are different
microRNA-adapted shRNAs compared to simple hairpin shRNAs
SMARTvector lentiviral shRNA particles in two cell lines
The SMARTvector no reporter option provides equivalent gene knockdown compared to the SMARTvector constructs with a fluorescent reporter