使用编码多个RNA干扰序列的增强型绿色荧光蛋白内含子剪接系统抑制端粒酶活性
Inhibition of Telomerase Activity Using an EGFP-Intron Splicing System Encoding Multiple RNAi Sequences.
作者信息
Sakiragaoglu O, Munn A L
机构信息
School of Medical Science and Menzies Health Institute Queensland, Griffith University (Gold Coast Campus), Parklands Drive, Southport, QLD, 4222, Australia.
出版信息
Mol Biotechnol. 2016 Dec;58(12):832-837. doi: 10.1007/s12033-016-9982-6.
To inhibit telomerase activity, a construct which contains artificial introns in the enhanced green fluorescent protein (EGFP) gene that encodes small hairpin RNA (shRNA) sequences that target human telomerase reverse transcriptase (hTERT) gene expression was designed and tested for its effect on lung cancer cell line. On intron splicing from the construct, intronic sequences were released and formed shRNA in the cells. After transfection of the construct, hTERT mRNA expression decreased by approximately 55 % in A549 cells. Correspondingly, in the same cell line, telomerase activity was decreased by approximately 23 %. The telomerase activity was transiently inhibited by this non-viral shRNA expression system that uses intron splicing to release artificial introns in an EGFP marker gene that contain shRNA targeting telomerase.
为抑制端粒酶活性，设计了一种构建体，该构建体在增强型绿色荧光蛋白（EGFP）基因中含有人工内含子，该基因编码靶向人端粒酶逆转录酶（hTERT）基因表达的小发夹RNA（shRNA）序列，并测试其对肺癌细胞系的影响。在构建体的内含子剪接过程中，内含子序列被释放并在细胞中形成shRNA。转染该构建体后，A549细胞中hTERT mRNA表达下降约55%。相应地，在同一细胞系中，端粒酶活性下降约23%。这种非病毒shRNA表达系统通过内含子剪接在含有靶向端粒酶的shRNA的EGFP标记基因中释放人工内含子，从而短暂抑制端粒酶活性。
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