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microDOWN™ miRNA Antagomir

Antagomir is chemically-modified single-strand miRNA inhibitor functioning by blocking miRNA regulation of target gene expression efficiently. They are synthesized to reduce the ability of endogenous miRNAs to silence target mRNA transcripts. They can down-regulate the corresponding endogenous miRNAs by either local or systemic injection into the animals.

Creative Biogene's microDOWN™ miRNA antagomir is single-strand miRNA inhibitor carrying the chemically modifications functioning by blocking miRNA regulation of target gene expression efficiently. The strand of the antagomir has 2 phosphorothioates at the 5' end, 4 phosphorothioates, 1 cholesterol group at the 3' end, and full-length nucleotide 2'-methoxy modification. Stability of microDOWN™ miRNA antagomir appears to be significantly higher than miRNA inhibitors. It exhibits enhanced cellular uptake, stability and regulatory activity and is recommended for miRNA functional studies in vitro and in vivo.

Figure 1. Mechanism of microDOWN™ miRNA antagomir.

Figure 2. qRT-PCR and western blot assays show the mRNA (A) and protein (B) expressions of TOB1 after transfection with antagomir-NC and antagomir-25.

Our microDOWN™ miRNA antagomir has unparalleled advantages

  1. Cover all human, mouse, rat miRNAs
  2. Optimized modifications for stable utility and easy transfection
  3. Long-term (up to 6 weeks) inhibition of the corresponding endogenous miRNAs
  4. No need of virus or vector for delivering
  5. Ready for in vivo and in vitro studies
  6. Various scales and grades for your application
  7. microDOWN™ antagomir negative control is available

Resuspension Protocol

  1. Briefly centrifuge tubes containing miRNA antagomir to ensure that the miRNA pellet is located at the bottom of the tube.
  2. Dissolve miRNA antagomir to a convenient stock concentration using the recommended volume of DEPC H2O (or RNase-free water). For example: Dissolve 1 OD miRNA antagomir to 20 µM using 250 µl DEPC H2O (or RNase-free water).
  3. Pipette the solution up and down 3-5 times (or vortex briefly).
  4. Briefly centrifuge tubes containing miRNA antagomir to ensure that the solution is collected at the bottom of the tube.
  5. Aliquot the miRNA antagomir into small volumes and store at -20°C. miRNA antagomir is stable (for 6 months under the specified storage condition). For best results, use in 3 months and limit freeze-thaw events for each tube no more than five times.

Cell Assay

Recommended amounts of microDOWN™ antagomir for your cell assay.

ReferencesmiRNACellAntagomir
Shan Y, et al. Gastroenterology. 2007miR-122Huh-7 WT, CNS350 nM
Brian D, et al. Mol Ther. 2007miR-206HeLa, MDA-231, MCF-720-50 nM
Zhu H, et al. Biochem Pharmacol. 2008miR-27a, miR-451A2780100 nM
Alla Musiyenko, et al. J Mol Med. 2008miR-126, miR-126LNCaP10~30 nM
Jia Yu, et al. PNAS. 2008miR-184, miR-205HeLa, HEKs1 μM
Federica et al. Cancer Res. 2008miR-221, miR-222Me665/150~250 nM
Relja Popovic, et al. Blood. 2009miR-196bbone marrow cells100 nM
Pascal Pineau, et al. PNAS. 2010miR-221, miR-222hepatocellular carcinoma-derived100 μM
Xiaogang Wang, et al. Nat Med. 2012miR-214MC3T3-E1, hFOB200 μM
Wang X, et al. Nat Med. 2013miR-214MC3T3-E1200 nM


In Vivo Delivery

ReferencesmiRNAAnimalDeliveryDosesVolumeInjection TimesDetection
Krützfeldt J, et al. Nature. 2005miR-122
miR-192
6-week-old micetail-vein injections80 mg per kg body weight0.2 ml per injection1 to 3 consecutive days24 h after the last injection
Jan Krützfeldt, et al. Nucleic Acids Research. 2007miR-166-week-old miceIntracerebral injection~800 ng per mouse0.2 ml per injectiona single injection72 h after injection
Alessandra Care, et al. Nature Medicine. 2007miR-1338-week-old miceminipump implantation80 mg per kg body weightN/Aa single infusion1 month after minipump implantation
Laura Fontana, et al. PLoS ONE. 2008miR-17-5p6-week-old nude miceintratumorally injection0.2 mg per mouse0.1 mlthree times per week for two weeks2 weeks after the first injection
Thomas Thum, et al. Nature. 2008miR- 2110~12 weeks old micejugular vein catheter80 mg per kg body weightN/Ainjected daily for three days3 weeks after the first injection
Neri Mercatelli, et al. PLoS ONE. 2008miR-221
miR-222
6-week-old miceintratumorally injection1 ug of each antagomir40 μlat day 0, 5 and 9, for a injections per tumorN/A
Ashuthosh Dharap, et al. J. Cereb. Blood Flow Metab. 2009miR-145ratminipump implantationtotally 960 pmol each rat48 μlN/AN/A
Li H, Xie H, et al. J Clin Invest. 2009miR-28616-week-old female micetail vein injection80 mg/kg body weight0.2 ml per injectiondays 1 to 3 for 3 consecutive weeks4 days, 3 weeks, and 6 weeks after the last injection
Bonauer A, et al. Science. 2009miR-92miceintravenous injection8 mg per kg of body weight0.2 ml per injectionat days 0, 2, 4, 7, 9 of the left coronary artery14 days
Jie Ding, et al. Nature cell biology. 2010miR-151-5pnude micetransplantion2x106 cells transfected by antagomirN/A5 weeks after injection
LinHui Liang, et al. Hepatology. 2010miR-125b6~8 weeks old nude micesubcutaneously injection2x105 cells transfected by antagomirN/A4 weeks after injection
Minfeng Shu, et al. Molecular Cancer. 2011miR-3354-week-old nude miceIntratumorally injection4 mg/ml50 μlevery two days for 2 weeks2 weeks after the first injection

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