Matched SMARTvector Inducible Negative and Positive RNAi Controls enable accurate data interpretation for both SMARTvector Inducible shRNA and shMIMIC Inducible Lentiviral microRNA
The SMARTvector Inducible RNAi Controls are cost-effective tools for determining optimal conditions for delivery and doxycycline induction, prior to using gene-specific SMARTvector Inducible Lentiviral shRNAs and shMIMIC Inducible Lentiviral microRNAs.
Start by identifying the optimal vector configuration for your cells with the SMARTchoice Inducible Non-targeting Control 4-Pack
Determine the most active promoter in the specific cells of interest by evaluating all four vectors simultaneously using the SMARTchoice Inducible Non-targeting Control 4-Pack (catalog # VSC6847):
- Four aliquots (25 µL each) of SMARTvector Inducible Non-targeting Controls supplied as high-titer, concentrated and purified lentiviral particles
- Includes each SMARTvector Inducible Non-targeting Control with PGK, mouse CMV, human EF1α and mouse EF1α promoters; and with the turboGFP fluorescent reporter
- Enables qualitative, empirical evaluation of optimal promoter driving expression of puromycin resistance and Tet-On 3G transactivator protein
Order SMARTvector inducible RNAi controls with promoter and reporter selections matching your gene-specific constructs.
SMARTvector Inducible vector-matched controls are available with options for all four promoters and two fluorescent reporters or without a fluorescent reporter. All individual controls are available as glycerol stocks and as 50 µL (2 x 25 µL) at 1 x 107 TU/mL, ± 20%, of purified, concentrated packaged lentiviral particles.
SMARTvector Inducible Non-targeting Controls
- Designed so that no known gene in human, mouse or rat is targeted, providing a baseline to which gene-specific effects can be normalized
- Useful for assessing transduction efficiency and the effects of transduction on the cells of interest
- SMARTvector Inducible Non-targeting Control 1: Our longest standing and most commonly used SMARTvector non-targeting control, ideal for supporting most experiments involving only a few genes
- SMARTvector Inducible Non-targeting Control 2 – 12: Designed specifically with the needs of screeners in mind, who may require using several different non-targeting control sequences in follow-up experiments; should be selected if multiple controls are desired
SMARTvector Inducible Positive Controls
- Permit confirmation of transduction efficiency and gene knockdown through consistent, potent silencing of non-essential, abundantly expressed housekeeping genes
- SMARTvector Inducible GAPD Positive Control shRNA (human, mouse and rat); validated to target glyceraldehyde-3-phosphate dehydrogenase (also known as GAPDH)
- SMARTvector Inducible PPIB Positive Control shRNA (human, mouse and rat); validated to target peptidylprolyl isomerase B (also known as cyclophilin B)
SMARTvector Inducible Non-targeting Control shRNA
SMARTvector Inducible RNAi Experimental Control
|Shipping Condition||Dry Ice, Frozen Gel Packs|
|Storage Conditions||-80 C|
|Stability at Recommended Storage Conditions||At least 12 months|
SMARTvector Inducible Lentiviral shRNA Experimental Workflow
Elements of the SMARTvector Inducible Lentiviral shRNA Backbone
|5' LTR||5' Long Terminal Repeat is necessary for lentiviral particle production and integration of the construct into the host cell genome|
|Ψ||Psi packaging sequence allows lentiviral genome packaging using lentiviral packaging systems|
|RRE||Rev Response Element enhances titer by increasing packaging efficiency of full-length lentiviral genomes|
|TRE3G||Inducible promoter with Tetracycline Response Elements which is activated by the Tet-On 3G protein in the presence of doxycycline|
|tGFP or tRFP||TurboGFP or TurboRFP reporter for visual tracking of transduction and expression upon doxycycline induction|
|SMARTvector universal scaffold||Optimized proprietary scaffold based on native primary microRNA in which gene-targeting sequence is embedded|
|PuroR||Puromycin resistance gene permits antibiotic selection of transduced cells|
|2A||Self-cleaving peptide that enables the expression of both PuroR and Tet-On 3G transactivator from a single RNA pol II promoter|
|Tet-On 3G||Encodes the doxycycline-regulated transactivator protein, which binds to TRE3G only in the presence of doxycycline|
|WPRE||Woodchuck Hepatitis Post-transcriptional Regulatory Element enhances transgene expression in target cells|
|3' SIN LTR||3' Self-inactivating Long Terminal Repeat for generation of replication-incompetent lentiviral particles|
SMARTvector Inducible shRNA promoter activities differ across cell types.
SMARTvector Inducible shRNAs enable tightly controlled knockdown of essential genes in a time- and doxycycline dose-dependent manner.