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; font-family: 等线; font-size: 9pt;">片特点:
• 覆盖度广—囊括了UniProt与Modomics中所有已知的tRNA修饰酶/蛋白因子
• 严谨性强—所有引物都通过了多样本严格验证
• 快速简易—即用型384孔板规格,只需将cDNA与qPCR Master Mix混合试剂加入PCR板中,4小时内得到实验结果。cDNA无需预先扩增。

 
聚焦tRNA修饰相关蛋白:
      Arraystar一直关注科学研究动态,设计开发市场需求的产品。近年来,tRNA修饰受到了越来越多的关注,tRNA修饰酶在多种疾病中的作用被陆续报道。在此背景下,Arraystar公司开发了tRNA修饰相关蛋白PCR芯片。此款芯片聚焦于tRNA转录后修饰过程中,发挥重要功能的酶及蛋白因子。Arraystar研究人员收录了多篇专业文献中的tRNA修饰相关蛋白[2, 5, 6],并汇总了UniProt和Modomics数据库中的相关酶与蛋白因子(表1)。为了保证数据质量,芯片包含了一系列参照,用于***DNA污染,反转录与qPCR的效率,亦或用于后续的数据标准化分析。
 
表1. tRNA修饰酶/蛋白因子(修饰类型)

tRNA修饰及相关蛋白 (85):

ADAT1(m1I37), ADAT2(m1I34), ADAT3(-), ALKBH1(mcm5U), ALKBH8(mchm5U34/mcm5s2U34/mcm5U34/mcm5Um34), C9orf64(-), CDK5RAP1(ms2A37/ms2 i6A37), CDKAL1(ms2A37/ms2 i6A37), CDKL1(ms2t6A), CTU1(mcm5S2U ), CTU2(mcm5S2U), DUS1L(D16/17), DUS2(D20), DUS3L(D47), DUS4L(D), ELP3(-), ELP4(mcm5s2U), FBLL1(-), FTSJ1(Cm32/Gm34), GTPBP3(tm5U34), HSD17B10(m1A9/m1G9), IKBKAP(-), KIAA0391(m1A9/m1G9), KIAA1456(Um), LAGE3(t6A37), LCMT2(o2yW), METTL1(m7G46), METTL2A(Cm), METTL2B(3mC), MOCS3(mcm5S2U ), MTO1(tm5U34), NAT10(Ac4C), NFS1(s2U34/tm5s2U34), NSUN2(m5C49), NSUN6(m5C72), OSGEP(t6A37), OSGEPL1(t6A37), PUS1(pseudouridine27/pseudouridine28), PUS10(pseudouridine 55), PUS3(pseudouridine39), PUS7L(pseudouridine), PUSL1(pseudouridine), QTRT1 (Q34), QTRT2 (Q34), RPUSD1(pseudouridine), RPUSD2(pseudouridine31/pseudouridine32), RPUSD3(pseudouridine), RPUSD4(pseudouridine31/pseudouridine32), SSB(-), TARBP1(-), THUMPD1(Acetylcytidine), THUMPD2(Gm), THUMPD3(Gm), TP53RK(t6A37), TPRKB(t6A37), TRDMT1(m5C38), TRIT1(i6A37), TRMO(m6t6A), TRMT1(m2G26/m22G26), TRMT10A(m1G9), TRMT10B(m1G9), TRMT10C(m1A9/m1G9), TRMT11(m2G10), TRMT112(mchm5U34/mcm5s2U34/mcm5U34/mcm5Um34), TRMT12(o2yW), TRMT13(m4C/m4A ), TRMT1L(m2G26/m22G26), TRMT2A(Um), TRMT2B(m5U54), TRMT44(Um44), TRMT5(m1G37/o2yW), TRMT6(m1A58), TRMT61A(m1A58), TRMT61B(m1A58), TRMU(s2U34/tm5s2U34), TRUB1(pseudouridine), TRUB2(pseudouridine55), TYW1(o2yW), TYW1B(o2yW), TYW3(o2yW), TYW5 (o2yW), UBA5(cyclic t6A), URM1(mcm5S2U), WDR4(m7G46), YRDC(t6A37)

tRNA修饰缩写参考Modomics数据库基因名参考UniProt

 

实验流程:

适用实时荧光定量PCR仪:
ABI ViiA™ 7, ABI 7500 & ABI 7500 FAST, ABI 7900HT, ABI QuantStudio™ 6 Flex Real-Time PCR system, ABI QuantStudio™ 7 Flex Real-Time PCR system, ABI QuantStudio™ 12K Flex Real-Time PCR System, Bio-Rad CFX384, Bio-Rad iCycler & iQ Real-Time PCR Systems, Eppendorf Realplex, QIAGEN Rotor Gene Q 100, Roche Light Cycler 480, Stratagene Mx3000, Roche Light Cycler 480

参考文献
[1] Kirchner S, Ignatova Z. Emerging roles of tRNA in adaptive translation, signalling dynamics and disease. Nature reviews Genetics 2015;16:98-112.
[2] El Yacoubi B, Bailly M, de Crecy-Lagard V. Biosynthesis and function of posttranscriptional modifications of transfer RNAs. Annual review of genetics 2012;46:69-95.
[3] Zhou B, Wei FY, Kanai N, Fujimura A, Kaitsuka T, Tomizawa K. Identification of a splicing variant that regulates type 2 diabetes risk factor CDKAL1 level by a coding-independent mechanism in human. Human molecular genetics 2014;23:4639-50.
[4] Zhang X, Cozen AE, Liu Y, Chen Q, Lowe TM. Small RNA Modifications: Integral to Function and Disease. Trends in molecular medicine 2016.
[5] Suzuki T, Nagao A, Suzuki T. Human mitochondrial tRNAs: biogenesis, function, structural aspects, and diseases. Annual review of genetics 2011;45:299-329.
[6] Torres AG, Batlle E, Ribas de Pouplana L. Role of tRNA modifications in human diseases. Trends in molecular medicine 2014;20:306-14.

 

 

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