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 miRNA前体(miRNA precusor )


     Biomics采用先进工艺合成与体内完全一致的高质量pre-miRNA,而且可以做到量产,突破了目前RNA合成序列长度只能在50碱基左右的障碍,大大提高了合成效率。将此合成的miRNA前体转染至细胞后,可形成与内源性成熟miRNA化学结构及生物功能一致的功能性单链。miRNA mimics是模拟pre-RNA人工设计的双链小RNA,虽然其化学结构和生物活性与内源性miRNA接近,但仍存在诸多差别。我们在研究中发现同样数量的pre-miRNA和miRNA mimics同时转染细胞时,pre-miRNA能更特异且高效的形成mature miRNA。还有研究报道,与双链的siRNA、shRNA、miRNA mimic相比,pre-miRNA的特殊结构对细胞、动物体内的非靶基因或功能影响更小,且干扰素效应更低,因此pre-miRNA在系统多因素的研究中具备独特的优势。

         增补miRNA表达活性以取得功能效应的研究
         筛选调控基因表达和影响细胞途径的miRNA
         鉴别验证miRNA靶点

产品名 规格 价格 周期

miRNA precusor

2.5 nmol

1000

2周

miRNA precusor

5nmol

1600

2周

miRNA precusor

10nmol

2500

2周

miRNA precusor control 2.5nmol 400 2周
Customer - defined miRNA precusor 5nmol 询价 2周

 
                   注:
                   1)Control可提供人,大鼠,小鼠现货,其他种属需定制
                   2)其他规格请询价
                   3)非常规种属miRNA precusor订购5nmol,起订量需达到5种以上


          纯度
               Biomics合成的pre-miRNA纯度高于85%

 
  HPLC分析pre-miRNA-21纯度为93.5%
   
 
  15%聚丙烯酰胺凝胶电泳
Lane1:ssRNA(47nt)
Lane2:ssRNA(47nt)
Lane3:pre-miRNA-21(75nt)


         稳定性
               Biomics合成的Pre-miRNA建议在-20℃或-70℃下存贮,在室温下可保持一个月左右

 
  15%聚丙烯酰胺变性胶检测pre-miRNA稳定性
Lane1:-70℃
Lane2:-20℃
Lane3:4℃
Lane3:室温(RT)
10nM的pre-miRNA溶液在-70℃,-20℃,4℃及室温下储存一个月,15%聚丙烯酰胺凝胶电泳检测RNA降解。


[1].Yoshinobu Shiba,et al,Chemical synthesis of a very long oligoribonucleotide with 2-cyanoethoxymethyl (CEM) as the 2’-O-protecting group: structural identification and biological activity of a synthetic 110mer precursor-microRNA candidate Nucleic Acids Research, 2007, Vol. 35, No. 10 3287–3296.
[2].David Brown et al,Metheds and compositions involving miRNA and miRNA inhibitor molecules,patent:US 8,058,250.
[3].Daniela Castanotto et al, Combinatorial delivery of small interfering RNAs reduces RNAi efficacy by selective incorporation into RISC Nucleic Acids Research, 2007, (35), 5154–5164.
[4].Kazuya Terasawa et al,Synthetic Pre-miRNA-Based shRNA as Potent RNAi Triggers Journal of Nucleic Acids (2011), 1-6。
[5].Dirk Grimm et al, Fatality in mice due to oversaturation of cellular microRNA/short hairpin RNA pathways Nature 2006( 441),537-541.
[6].Richard I. Gregory,et al,Human RISC Couples MicroRNA Biogenesis and Posttranscriptional Gene Silencing Cell, Vol. 123, 631–640, November 18, 2005.
[7].M Bauer et al,Prevention of interferon-stimulated gene expression using microRNA-designed hairpins Gene Therapy (2009) 16, 142–147.
[8]. Onsam Sin et al,Gene Silencing Efficiency and INF-b Induction Effects of Splicing miRNA 155-Based Artificial miRNA with Pre-miRNA Stem-Loop Structures, Biochem Genet (2012) 50:112–121.
[9]. Carol A. Sledz et al Activation of the interferon system by short-interfering RNA, Nature Cell Biology 2003(9) 834-839.


 

 

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