tRNA测序服务

      转运RNA（tRNA）是一类广泛存在于生物体内，在肽链合成中负责氨基酸转运的非编码小RNA分子。研究表明，tRNA除参与肽链合成外，还参与多种生理病理学过程[1]。不同机体组织与细胞的转录组存在明显差异，且不同mRNA的密码子使用偏好性不同，使得所需的tRNA库存在显著差异。tRNA库与转录组密码子的匹配度影响着翻译效率和准确性[2-4]。增殖[5]、分化[5, 6]和凋亡[7]等细胞生理状态都受到tRNA库变化的影响。再者，许多研究表明tRNA库在肿瘤[5, 8-12]、Ⅱ型糖尿病[13]、亨廷顿舞蹈症[14]以及病毒感染[15]等疾病的发生发展中起着重要的调控作用。全长tRNA分子具有非常稳定的二级结构和高水平的核苷酸修饰（tRNA功能发挥所必须的），这些都会严重影响高通量测序文库构建过程反转录酶的全长延伸活性以及核酸序列的比对分析，从而导致常规small RNA建库方案针对tRNA分子的文库构建失败及分析结果的不准确。
      Aksomics（原康成生物）在现有新型tRNA测序方案Hydro-tRNAseq (Cell Reports,2017)的基础上进行了进一步的优化，添加修饰核苷酸的去甲基化处理流程，更为优势地克服了tRNA测序的各个难点，高检出率且准确定量各类tRNA分子（图1）。
 

 

 

 

 

tRNA测序服务优势
● 特异性的tRNA测序文库构建方案，高tRNA检出率及定量准确性
● 优化的tRNA去甲基化修饰流程，进一步提升tRNA检测分析效率
● 全面的tRNA转录组参考库，综合了所有的权威数据库
● 详尽的tRNA注释信息与生物信息学分析
● 可直接用于文章发表的高质量图片
● 无缝链接康成生物提供的其它tRNA分析服务平台，包括tRF&tiRNA-seq, tRF&tiRNA PCR array, tRNA Repertoire PCR array, tRNA Modification Enzymes PCR Array与LC-MS based tRNA Mod-ification analysis.
 

基础数据分析结果展示
1. 基本数据分析结果展示
1.1 tRNA表达定量
      康成生物利用Arraystar独有的tRNA数据分析流程，提供信息全面的数据集（图2）。除各tRNA表达量外，还额外提供有tRNA二级结构及密码子位置等相关信息，更好地为客户后续科研助力。

图2：全面的tRNA测序结果数据集

1.2 tRNA差异表达分析
      分析tRNA表达谱，挖掘差异表达tRNA分子；通过Heatmap、Scatter Diagram、Volcano Plot等统计图形能够清晰明确的展示样本之间的tRNA表达差异性（图3）。

图3：tRNA差异表达Scatter Diagram和Volcano Plot示例。
 

参考文献
[1] Giege R. Toward a more complete view of tRNA biology. Nat Struct Mol Biol 15, 1007-14. (2008), PMID:18836497.
[2] Drummond DA, Wilke CO. Mistranslation-induced protein misfolding as a dominant constraint on coding-sequence evolution. Cell 134, 341-52. (2008), PMID:18662548.
[3] Plotkin JB, Kudla G. Synonymous but not the same: the causes and consequences of codon bias. Nat Rev Genet 12, 32-42. (2011), PMID:21102527.
[4] Shah P, Gilchrist MA. Explaining complex codon usage patterns with selection for translational efficiency, mutation bias, and genetic drift. Proc Natl Acad Sci U S A 108, 10231-6. (2011), PMID:21646514.
[5] Gingold H, et al. A dual program for translation regulation in cellular proliferation and differentiation. Cell 158, 1281-92. (2014), PMID:25215487.
[6] Pavon-Eternod M, et al. Overexpression of initiator methionine tRNA leads to global reprogramming of tRNA expression and increased proliferation in human epithelial cells. RNA 19, 461-6. (2013), PMID:23431330.
[7] Mei Y, et al. Apoptotic regulation and tRNA. Protein Cell 1, 795-801. (2010), PMID:21113408.
[8] Goodarzi H, et al. Modulated Expression of Specific tRNAs Drives Gene Expression and Cancer Progression. Cell 165, 1416-27. (2016), PMID:27259150.
[9] Berns A. A tRNA with oncogenic capacity. Cell 133, 29-30. (2008), PMID:18394985.
[10] Waldman YY, et al. TP53 cancerous mutations exhibit selection for translation efficiency. Cancer Res 69, 8807-13. (2009), PMID:19887606.
[11] Kushner JP, et al. Elevated methionine-tRNA synthetase activity in human colon cancer. Proc Soc Exp Biol Med 153, 273-6. (1976), PMID:995958.
[12] Marshall L, et al. Elevated tRNA(iMet) synthesis can drive cell proliferation and oncogenic transformation. Cell 133, 78-89. (2008), PMID:18394991.
[13] Krokowski D, et al. A self-defeating anabolic program leads to beta-cell apoptosis in endoplasmic reticulum stress-induced diabetes via regulation of amino acid flux. J Biol Chem 288, 17202-13. (2013), PMID:23645676.
[14] Girstmair H, et al. Depletion of cognate charged transfer RNA causes translational frameshifting within the expanded CAG stretch in huntingtin. Cell Rep 3, 148-59. (2013), PMID:23352662.
[15] van Weringh A, et al. HIV-1 modulates the tRNA pool to improve translation efficiency. Mol Biol Evol 28, 1827-34. (2011), PMID:21216840.
[16] Gogakos T., et al. Characterizing expression and processing of precursor and mature human tRNA by Hydo-tRNAseq and RAP-CLIP. Cell reports 20(6):1463-1475. (2017), PMID:28793268