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3种人工核酸酶的原理及其应用探析(2)

来源:学术堂 作者:朱老师
发布于:2017-04-12 共8478字
  同年,我们[33]与合作者又利用CRISPR/Cas9技术对食蟹猴基因组进行了修饰,获得多位点基因敲除的双胞胎食蟹猴,这是CRISPR/Cas9技术首次应用于非人灵长类,这对于构建一些由多基因控制的复杂疾病( 例如帕金森氏病) 的动物模型的研究有非常重要的意义。通过检测新生小猴脐带和胎盘的DNA样本,没有发现脱靶现象。此外,我们利用单细胞测序技术对流产胎儿的生殖系细胞检测后,发现生 殖 细 胞 也 存 在 基 因 突 变,首 次 证 明 了 利 用CRISPR / Cas9技术介导的基因突变可以实现生殖系传递[42].
  
  3. 3基因治疗
  
  基因治疗是利用基因编辑技术将表达异常的致病基因纠正为正常基因,从而实现治疗疾病的目的。传统的基因治疗难以将细胞中有缺陷的基因精准替换成正常基因片段,而且可能会在细胞中残留其它元件对机体产生很大的毒副作用。而基因编辑新技术则可以精确定位靶基因,在靶位点对基因实施编辑或进行切除,从而达到基因治疗的目的。
  
  Li等[43]利用ZFN技术在小鼠体内成功治愈了血友病,该研究首先敲除突变的基因,然后再利用病毒载体将正常的基因导入宿主。传统的基因编辑技术很难实现对线粒体遗传信息的改造,而Bacman等[44]利用TALEN技术成功敲除了线粒体内的突变基因。这是TALEN技术首次应用于线粒体基因的编辑,这项研究有望用于治疗由线粒体DNA突变引起的疾病。
  
  CRISPR / cas9系统作为第3代人工核酸酶技术凭借其独特的优势,在基因治疗方面有着更加广泛的应用。 Schwank等[45]利用CRISPR/cas9技术纠正了与囊性纤维化有关的缺陷基因;Hu等[46]利用CRISPR / cas9技术首次成功地把艾滋病病毒从人类细胞中清除,这将极大的推动治愈艾滋病的进程;Yin等[47]利用CRISPR/cas9技术将突变可导致酪氨酸血症Ⅰ型的FAH基因修饰,约成功纠正了部分肝细胞的基因突变,使正常的肝细胞细胞恢复至占肝细胞总量1 /3,这足以治愈该病。
  
  4问题与展望
  
  人工核酸酶介导的基因编辑技术凭借自身独特的优势,极大的推动了生命科学研究的进程。但是,即使是TALEN、CRISPR/cas9这些能够精确进行基因修饰的人工核酸酶技术也处于发展的初级阶段,研究者需要根据不同的实验需求选择合适的人工核酸酶技术进行基因组修饰。此外,体细胞核移植技术是获得转基因动物最为可靠和有效的方法。在未来的研究中,将人工核酸酶技术与核移植技结合,可以更加高效地建立人类疾病的动物模型,为科学家们深入研究疾病发病机制和探索新的疾病治疗方案提供更多可能。
  
  参考文献(References)
  
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