shRNA Cloning Vectors

This product line features cloning of specific hairpin sequences into one of our standard or customized shRNA lentiviral expression vectors; or we will design and clone 3 to 5 shRNA constructs targeting any human or mouse gene.

Following cloning, constructs are sequenced and we provide you with lentiviral plasmid DNA and, optionally, pre-packaged, ready-to-transduce lentiviral particles.

Our lentiviral service is typically used by researchers who have found hits using a Cellecta pooled shRNA library, or who are interested in knocking down a specific gene. You can provide us with the hairpin sequences you want to clone, the reference for particular shRNAs in one of our libraries, or just ask us to design a few constructs to your target of interest. We offer a range of vectors with different fluorescent and selection markers with which to make constructs. We will synthesize the DNA to encode the desired hairpin, clone it, and sequence it to verify correct construction. Optionally, we offer packaging services to produce lentiviral particles that may be used to directly transduce your cells of interest.

Overall, about 70% of our constructs knock down the target by more than 70% based on qRT-PCR, so it is likely that at least 2 of 3 or 3 of 5 constructs will be effective. However, we do not guarantee this level for any specific target, and the percentage of highly effective shRNAs will vary from target to target. Also, you may see a phenotype with just a small percent knockdown or require a much higher knockdown before a phenotype manifests itself, depending on the nature of the gene target. If you choose to have us design the sgRNA sequences, you will receive several of our best-designed constructs to test and characterize so you can determine which of these best match your experimental needs.

Citations:

• Wilms tumorigenesis in human kidney organoids (2021)
• Paracrine placental growth factor signaling in response to ionizing radiation Is p53-dependent and contributes to radioresistance (2021)
• Proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), play an oncogenic role in chronic myeloid leukemia by stabilizing nuclear factor-kappa B (2021)
• USP19 deubiquitinates EWS-FLI1 to regulate Ewing sarcoma growth (2019)
• ZEB1/NuRD complex suppresses TBC1D2b to stimulate E-cadherin internalization and promote metastasis in lung cancer (2019)