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Research Article

RNA靶向CRISPR-Cas13a有效下调BCR-ABL致瘤性

卷 21, 期 3, 2021

发表于: 17 February, 2021

页: [270 - 277] 页: 8

弟呕挨: 10.2174/1566523221666210217155233

价格: $65

摘要

目标：利用CRISPR Cas13a诱导BCR-ABL基因沉默。 背景：CML是一种克隆性骨髓增生性多能干细胞疾病，由染色体9和22之间的相互易位驱动，形成BCR-ABL融合基因。像伊马替尼这样的酪氨酸激酶抑制剂药物是治疗的主要药物，如果没有相容的干细胞供体，对这些药物有耐药性的病例预后较差。然而，随着基因编辑技术的快速发展，目前大多数研究都集中在开发一种针对单基因疾病的翻译模型，具有潜在的永久治愈。 目的:探讨RNA靶向CRISPR-Cas13a系统在CML细胞系K562中有效敲除BCR-ABL融合转录本的潜在应用价值。 方法：设计特异于嵌合BCR-ABL基因的CRISPR Cas13a crRNA，并将该系统作为双质粒系统转染到CML细胞系K562中。通过评估依赖于BCR-ABL基因表达的下游基因的表达水平来列举影响。此外，下一代测序也被用来确定CRISPR对该基因的影响。 结果：CRISPR系统成功降低了依赖于激活的BCR-ABL激酶信号的下游基因[pCRKL和pCRK]的表达高达4.3倍。crispr处理后的细胞活力也显著降低了373.83倍[p-value= 0.000891196]。时间依赖动力学也强调了显著的体外抑制活性，可持续长达8周[p-value: 0.025]。根据Oxford MinION下一代测序器的cDNA测序数据，CRISPR处理的细胞显示62.37%的疑似裂解读取。 结论：这些初步结果突出了RNA靶向CRISPRs在血液学肿瘤(如CML)中的极好的潜在应用，并为这一方向的进一步研究铺平了道路。

关键词: CRISPR，肿瘤学，分子生物学，血液学，RNA裂解，BCR-ABL

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