miR-Tuners Set of 3 Control vectors
|LV104||3 x 10 µg|
The Set of 3 Control vectors contains Low, Med or Hi miR-Tuner elements cloned within the 3’UTR of the reporter gene Firefly Luciferase. This allows for simple validation of the miR-Tuner system by measuring and comparing output luminescence. Luminescence can be quantified using abm’s Luciferase Assay Kit (Cat. No. G287).
miR-Tuners are short, synthetic miRNA response element sequences which reside in the 3’ UTR of the gene of interest (GOI). These sequences are responsive to a native miRNA species which is widely expressed in many human and mouse cells. Using the miR-Tuner Low, Med or Hi will allow for precise low, medium or high expression levels of the GOI, respectively.
Fine-tuning transgene expression levels in mammalian cells for:
•Engineered genetic circuits and cell lines
•Studying physiological and biochemical changes
|Unit quantity||3 x 10 µg|
- Lentiviral Vector Amplification Protocol
- Selection-Drug Killing Curve
- Lentivector Packaging Protocol
- Lentivirus Infection Protocol
- miR-Tuners Set of 3 Control vectors Datasheet
- Enhanced Lentivirus Safety Features: Replication Incompetency
- Suggested MOI for Common Cancer Cell Lines
- Lentiviral System FAQ
- Control Vectors and Viruses (Blank, GFP, Scrambled, and more)
|Is the 10 ml of quantity for lentivirus or vector? I'd like to get a lenti vector expresses both GFP and my gene of interest like neuroglobin.I will use the lenti vector with packaging vectors to make virus in 293T cells, and then to transduce my target cells with the virus. Can you make the vector for me?|
The 10ml is ready-to-use lentivirus. For your second question, we offer custom gene cloning at affordable prices. If you have your gene in your plasmid with a compatible site with our vector, the subcloning will be $450 per construct. An extra cost will be charged if we have to find the gene for you.
|What is the EGFP Lentivirus Map (cat#LV006)?|
It is cloned in a Lenti-His vector using EcoRI and XhoI cut sites. There is a stop codon at the end of the the EGFP gene so the tag is not expressed.
|What is the wave length the GFP gene?|
Our GFP has a excitation of 488 nm and emission of 507 nm.
|About the infection protocol, how many infections can I do with the 10ml preparation?|
Based on the instruction coming with the package, 2ml is need for a 6-well, thus the 10ml is only enough for 5 infections. This is likely to be enough for generating a stable cell line.
|If I transfect or infect with an EGFP construct, when will I begin to observe fluorescence?|
Enhanced green fluorescent protein (EGFP), often used to check transfection efficiency or viral titer, can be visualized in packaging cells 24-48 hours after transfection. In target cells, EGFP is usually detected 48-96 hours after infection, depending on the cell type and infection efficiency.
|What is the source for the luciferase gene on the Lenti-UBC-Luc Lentivirus (Cat#LV051)?|
It is from firefly. Luc2 emission spectrum peaks at 600nm in mammalian cells. The substrate is regular D-luciferin.
|How is the IRES-EGFP construct signal compare to the normal GFP control?|
EGFP signal is much weaker due to IRES elements. The levels of EGFP expression is effected by surrounding genetic elements regardless of the high viral particles.
|Can the expression of the lentiviral vector differ among different cellines (for eg,HeLa,MCF 7,293T etc)|
Yes, like transfection, lentiviral vector transduction efficiency is cell line dependent. 293T will have the highest transduction efficiency.
|Are the WPRE and cPPT elements present?|
Both elements are present in all of our pLenti-III vectors. The WPRE is located immediately after the Puromycin marker (between the Puromycin and the 3'LTR). The cPPT is located immediately before the promoter (UbC/PGK/EF1a), between the 5'LTR and the promoter.
|For Cat# LV152, is the 3'LTR region truncated?|
No, this is a 2nd generation construct and the 3'LTR region is not truncated.
|What is the source for the luciferase gene in the Lenti-PGK-Luc Lentivirus (Cat#LV088)?|
It is from firefly (Photinus pyralis)
|Does LV152 contain regular Luc or Luc2?|
LV152 contains Luc2
|What is the difference between Luc2 and regular Luc?|
Luc2 is an engineered firefly luciferase, which is engineered to remove most cryptic transcription factor binding site and improve mammalian expression.
- Manshouri, R., Coyaud, E., Kundu, S. T., Peng, D. H., Stratton, S. A., Alton, K., ... & Carugo, A. "ZEB1/NuRD complex suppresses TBC1D2b to stimulate E-cadherin internalization and promote metastasis in lung cancer" Nature communications 10(1):1-15 (2019).
- Nihalani, D., Solanki, A. K., Arif, E., Srivastava, P., Rahman, B., Zuo, X., … Lipschutz, J. H. "Disruption of the exocyst induces podocyte loss and dysfunction" Journal of Biological Chemistry 294(26):10104–10119 (2019). DOI: 10.1074/jbc.ra119.008362.
- Nischalke, H. D., Lutz, P., Bartok, E., Krämer, B., Langhans, B., Frizler, R., ... & Stickel, F. "The PNPLA3 I148M variant promotes lipid-induced hepatocyte secretion of CXC chemokines establishing a tumorigenic milieu" Journal of Molecular Medicine 1-12: (2019).
- Peruzzaro, S.T., Andrews, M.M.M., Al-Gharaibeh, A. "doi:10" 1186/s12974-018-1383-2 : (2019). DOI: 10.1186/s12974-018-1383-2.
- Rahnama, M. A., Movassaghpour, A. A., Soleimani, M., Atashi, A., Anbarlou, A., & Asenjan, K. S. "MicroRNA-15b target Sall4 and diminish in vitro UCB-derived HSCs expansion" EXCLI journal 14:601 (2015).
- Sarret, C., Ashkavand, Z., Paules, E., Dorboz, I., Pediaditakis, P., Sumner, S., … Krupenko, S. A. "Deleterious mutations in ALDH1L2 suggest a novel cause for neuro-ichthyotic syndrome" Npj Genomic Medicine 4(1): (2019). DOI: 10.1038/s41525-019-0092-9.
- Spörrer, M., Prochnicki, A., Tölle, R. C., Nyström, A., Esser, P. R., Homberg, M., … Kiritsi, D. "Treatment of keratinocytes with 4-phenylbutyrate in epidermolysis bullosa: Lessons for therapies in keratin disorders" EBioMedicine 44:502–515 (2019). DOI: 10.1016/j.ebiom.2019.04.062.
- Wang, . "U" S. Patent Application No. 0071651 A1 : (2019).
- Von Dwingelo, J., Chung, I. Y. W., Price, C. T., Li, L., Jones, S., Cygler, M., & Abu Kwaik, Y. "Interaction of the Ankyrin H Core Effector of Legionella with the Host LARP7 Component of the 7SK snRNP Complex" mBio 10(4): (2019). DOI: 10.1128/mbio.01942-19.
- Wang, H. H., Wu, Y. J., Tseng, Y. M., Su, C. H., Hsieh, C. L., & Yeh, H. I. "Mitochondrial fission protein 1 up-regulation ameliorates senescence-related endothelial dysfunction of human endothelial progenitor cells" Angiogenesis 1-14: (2019).
- Wang, Q., Yu, H., Yu, H., Ma, M., Ma, Y., & Li, R. "miR‑223‑3p/TIAL1 interaction is involved in the mechanisms associated with the neuroprotective effects of dexmedetomidine on hippocampal neuronal cells in�vitro" Molecular Medicine Reports. : (2018). DOI: 10.3892/mmr.2018.9742.
- Xu, T., He, B. S., Pan, B., Pan, Y. Q., Sun, H. L., Liu, X. X., ... & Wang, S. K. "MiR‐142‐3p functions as a tumor suppressor by targeting RAC1/PAK1 pathway in breast cancer" Journal of Cellular Physiology. : (2019).
- Zhang, B., Roosmalen, I. A. M., Reis, C. R., Setroikromo, R., & Quax, W. J. "Death receptor 5 is activated by fucosylation in colon cancer cells" The FEBS Journal 286(3):555–571 (2019). DOI: 10.1111/febs.14742.
- Zhang, L., Irimia, A., He, L., Landais, E., Rantalainen, K., Leaman, D. P., ... & Poignard, P. "An MPER antibody neutralizes HIV-1 using germline features shared among donors" Nature Communications 10(1):1-16 (2019).
- Zhang, R.. "N6-methylation of adenosine (m6A) of FZD10 mRNA contributes to PARP inhibitor resistance" AACR. Doi:10.1158/0008-5472 : (2019).
- Zhang, Y. L., Zhou, H. Y., Ma, Q. Z. "US Patent No" 20190135937A1. Retrieved from : (2018).
- Zhao, Q., Zhao, S., Li, J., Zhang, H., Qian, C., Wang, H., … Zhao, Y. "TCF7L2 activated HOXA-AS2 decreased the glucocorticoid sensitivity in acute lymphoblastic leukemia through regulating HOXA3/EGFR/Ras/Raf/MEK/ERK pathway" Biomedicine & Pharmacotherapy 109:1640–1649 (2019). DOI: 10.1016/j.biopha.2018.10.046.