Lenti-UTR-Dual-Luciferase Blank Virus
Control lentivirus expressing firefly luciferase and renilla luciferase.
|Unit quantity||3.0 ml|
1 year when stored at -20°C or lower in a non-frost free freezer.
- Lentiviral Vector Amplification Protocol
- Selection-Drug Killing Curve
- Lentivirus Packing Protocol (for use with Vectors only)
- Lentivirus Infection Protocol
- UTR Reporters Handbook
- Enhanced Lentivirus Safety Features: Replication Incompetency
- Suggested MOI for Common Cancer Cell Lines
|I am very interested in your miRNA products. How about the price for the 3' UTR Luciferase/GFP Reporters? Could I directly order with your company or online and pay by PO?|
To search for the price of 3'UTR product, please go to http://www.abmgood.com/microRNA-miRNA/targetvalid.php?csn=26&ssn=11838&dsn=12288, and enter the gene name. You can place your order by email, telephone, fax, mail or online. PO number or Visa can be used. Please see the detail by clicking the following link http://www.abmgood.com/misc/orders.php
|Is it possible to use your kit (miRNA cDNA Synthesis Kit) to do quantitative PCR with the Taqman strategy?|
This kit along with our miRNA primers would not be compatible for use with Taqman strategy.
|What is the source of the Luciferase gene?|
It is from the firefly.
|How does the 3'UTR platform work?|
The stable cell line or the infected cells from the lentivirus will consistently express luciferase/GFP unless there are miRNA that activate the 3'UTR to "knockdown" the reporter expression.
|What is the forward and reverse sequencing primers for the 3'UTR luciferase reporter systems?|
Forward sequencing primer: Luc Forward primer 5'-GCAAGTTGGACGCCCGCAAGATC-3' Reverse sequencing primer: SV40 promoter reverse primer 5`-TAGTCAGCCATGGGGCGGAGA -3'
|Whats the parental cell line for the 3'UTR stable cells?|
|What is the upper size limit for the 3'UTR insert for viral packaging?|
For GFP, we can fit a 3.5kb insert for efficient packaging. For Luc, we can fit a 2.6kb insert for efficient packaging. The above size limits can be stretched a little (~0.5kb-1kb larger), but the packaging efficiency and virus titer will be lower.
|How does the universal reverse primer work?|
The universal reverse primer can be used for all mature miRNA. To quantify or amplify a specific miRNA, the universal reverse primer with the appropriate forward miRNA primer needs to be used. Our forward primers can work with any kits. Our universal primers will only work with abm's cDNA synthesis kit.
|Can I use SYBR green mastermix for downstream qPCR of the cDNA produced with this kit?|
It may be possible to use a SYBR green mastermix for qPCR, however, our BrightGreen miRNA miRNA qPCR MasterMix recipe has been fully optimized to provide the most successful conditions for qPCR. We have performed extensive testing with minor differences in the composition of miRNA qPCR buffer, this has proven to give very dramatic differences to the results of qPCR. General qPCR mastermixes have more additives to prevent non-specificity and formation of primer dimers and the conditions are often too harsh for the miRNA primers to anneal properly
|Could 3'UTR GFP reporter be used with a conventional transient transfection system (e.s.Lipofectamin 2000)? Could this vector be used to validate the activity of an endogenous overexpressed miRNA?|
Yes, this vector can also be transiently transfected using a transfection reagent such as lipofectamine 2000, or DNAfectin available from abm. This reporter construct can also be used to validate interactions of an endogenously expressed miRNA, if the known target site is present in the 3' UTR sequence.
- Mazan-Mamczarz, K et al. "Down-Regulation of eIF4GII by miR-520c-3p Represses Diffuse Large B Cell Lymphoma Development " PLOS GENETICS 10 (1):e1004105 (2014). DOI: 10.1371/journal.pgen.1004105. PubMed: 24497838.
- Liang, Y., Song, X., Li, Y., Su, P., Han, D., Ma, T., … Yang, Q. "circKDM4C suppresses tumor progression and attenuates doxorubicin resistance by regulating miR-548p/PBLD axis in breast cancer" Oncogene 38(42):6850–6866 (2019). DOI: 10.1038/s41388-019-0926-z.
- Verma, M., Asakura, Y., & Asakura, A. "Inhibition of microRNA‐92a increases blood vessels and satellite cells in skeletal muscle but does not improve duchenne muscular dystrophy–related phenotype in mdx mice" Muscle & Nerve 59(5):594–602 (2019). DOI: 10.1002/mus.26433.
- Wu., Chien-Wei., . "Downregulation of MiR-144 by Triptolide Enhanced p85α−PTEN Complex Formation Causing S Phase Arrest of Human Nasopharyngeal Carcinoma Cells" European Journal of Pharmacology 855:137-148 (2019). DOI: 10.1016/j.ejphar.2019.04.052..
- Wu, M.-J., Chen, Y.-S., Kim, M. R., Chang, C.-C., Gampala, S., Zhang, Y., … Chang, C.-J. "Epithelial-Mesenchymal Transition Directs Stem Cell Polarity via Regulation of Mitofusin" Cell Metabolism 29(4):993–1002.e6 (2019). DOI: 10.1016/j.cmet.2018.11.004.
- Xiang, X., Zhou, Y., Sun, H., Tan, S., Lu, Z., Huang, L., & Wang, W. "Ivabradine abrogates TNF-α-induced degradation of articular cartilage matrix" International Immunopharmacology 66:347–353 (2019). DOI: 10.1016/j.intimp.2018.11.035.
- Zhou, Y., Lei, J., Xie, Q., Wu, L., Jin, S., Guo, B., ... & Zhang, J. "Fibrinogen-like protein 2 controls sepsis catabasis by interacting with resolvin Dp5" Science Advances 5(11):eaax0629 (2019).
- Zhu, R., Xue, X., Shen, M., Tsai, Y., Keng, P. C., Chen, Y., … Chen, Y. "NFκB and TNFα as individual key molecules associated with the cisplatin-resistance and radioresistance of lung cancer" Experimental Cell Research 374(1):181–188 (2019). DOI: 10.1016/j.yexcr.2018.11.022.