shRNA Knockdown Constructs
Cellecta provides complete services for construction of custom constructs expressing shRNA targeting any transcript.
- Choose constitutive or inducible version of H1 or U6 shRNA promoters
- Select GFP, RFP, PuroR, BleoR, NeoR, or Hygro-HK markers
- Obtain constructs as plasmid or packaged lentiviral particles
To make an shRNA construct, we only need to know the RefSeq number. We can design any number of shRNA constructs that target the transcript. Our proprietary shRNA design delivers approximately 70% shRNAs that generate >70% transcript knockdown in standard cell lines as measured by qRT-PCR.
We provide three types of shRNA plasmid construction services:
Simple shRNA Cloning: We can clone a specific shRNA into one of our standard shRNA expression vectors, or you can have us custom-design a vector for you. Often utilized by researchers who have found hits using a Cellecta pooled shRNA library, this service is very straightforward. You simply need to provide us with the shRNA sequence you want or reference an shRNA in one of our libraries, and we will make the insert, clone it, and sequence it to verify correct construction.
Targeted Sets of shRNA Constructs: We can provide 3 to 5 shRNA constructs targeting a specific human or mouse gene. These are constructed based on design parameters we use in creating our libraries. 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, depending on the characteristics of your target gene, the level of transcript knockdown may not accurately correspond to a phenotypic effect. In other words, you may see a phenotype with just a small percent knockdown or require a much higher knockdown before a phenotype manifests. With this service, you receive several of our best-designed constructs to test and characterize to see which best matches your experimental needs.
Validated shRNA Expression Construct: We can design and make a validated shRNA expression construct that is confirmed by qRT-PCR to knock down any desired target gene transcript by at least 70% in the cell line of your choice. For this service, we design and make several constructs expressing different shRNAs to the target gene (as is done in the Targeted Sets service described above), test knockdown by qRT-PCR, and provide the most effective construct.
- Honda T, Yamamoto Y, Daito T, Matsumoto Y, Makino A, Tomonaga K. Long-term expression of miRNA for RNA interference using a novel vector system based on a negative-strand RNA virus. Sci Rep. 2016 May 18;6:26154. doi: 10.1038/srep26154. PubMed PMID: 27189575. (used as miRNA vector)
- Zhou J, Wan J, Gao X, Zhang X, Jaffrey SR, Qian SB. Dynamic m6A mRNA methylation directs translational control of heat shock response. Nature. 2015 Oct 12. doi: 10.1038/nature15377. [Epub ahead of print] PubMed PMID: 26458103.
- Han Y, Gao X, Liu B, Wan J, Zhang X, Qian SB. (2014) Ribosome profiling reveals sequence-independent post-initiation pausing as a signature of translation. Cell Research. PMID: 24903108
- Khorashad JS, Mason CC, Kraft IL, Reynolds KR, Pomicter AD, Eiring AM, Zabriskie MS, Iovino AJ, Heaton W, Tantravahi SK, Kauffman M, Schacham S, Chenchik A, Bonneau K, O’Hare T, Deininger MW. (2013) An Unbiased shRNA Library Screen Identifies Nucleocytoplasmic Transport As a Potential Target For Treatment Of Chronic Myeloid Leukemia. Blood. 122(21):2707.
- Mele DA, Salmeron A, Ghosh S, Huang HR, Bryant BM, Lora JM. (2013). BET bromodomain inhibition suppresses TH17-mediated pathology. J Exp Med. Oct 21;210(11):2181-90. doi: 10.1084/jem.20130376. Epub 2013 Oct 7. PubMed PMID: 24101376; PubMed Central PMCID: PMC3804955.
- Miyazaki H, Higashimoto K, Yada Y, Endo TA, Sharif J, Komori T, Matsuda M, Koseki Y, Nakayama M, Soejima H, Handa H, Koseki H, Hirose S, Nishioka K. (2013) Ash1l methylates lys36 of histone h3 independently of transcriptional elongation to counteract polycomb silencing. PLoS Genetics.
To place an order or simply calculate pricing, please fill out the shRNA Construct Order Form and email it to us at firstname.lastname@example.org. Please indicate whether you are placing an order or requesting a quote. If placing an order, please refer to the PO number and email the PO to email@example.com or fax it to 650-938-3911.
For our selection of positive and negative shRNA control constructs, please see our Control Constructs web page.