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多重基因编辑以促进细胞存活使用低pH集群定期间隔的短palindromic重复激活 (CRISPRa) 基因扰动基因

Hunter Levis¹, Jacob Weston¹, Brooke Austin¹

¹Department of Biomedical Engineering, The University of Utah, Salt Lake City, Utah, USA.

|May 28, 2023

概括

这项研究使用CRISPR基因编辑来识别改善干细胞在酸性环境中的存活率的基因,这对于治疗退行性磁盘疾病 (DDD) 和其他疾病至关重要. 这些发现提高了细胞治疗在具有挑战性的微环境中的有效性.

科学领域:

生物技术是生物技术.
再生医学是一种再生医学.
遗传学遗传学是一个

背景情况:

腰部疼痛,通常是由退行性磁盘疾病 (DDD) 引起的,是全球领先的残疾.
目前的DDD治疗主要是缓解性,缺乏恢复组织功能的潜力.
细胞疗法为DDD提供了希望,但由于退化的磁盘的酸性微环境而面临挑战,这损害了干细胞的生存能力.

研究的目的:

通过使用CRISPR激活查,确定增强脂肪衍生干细胞在酸性条件下生存的基因标.
为了在DDD中改进潜在的治疗应用,设计具有提高活力的干细胞.

主要方法:

使用CRISPR激活 (a) 基因扰动屏幕来识别支持生存的基因.
在脂肪基因干细胞中通过细胞活力测试 (细胞计数套件-8) 验证候选基因.
在颗粒培养中评估了工程干细胞的细胞外矩阵生产.

主要成果:

确定了1213个潜在的亲生存基因,并将其缩小到20个用于验证.
选择并验证了前五个支持生存的基因.
证明了工程干细胞的细胞外基质生产能力.

结论:

克里斯普尔屏幕可以设计细胞表型,以提高干细胞在酸性环境中的生存能力.
这种方法有可能改善DDD和其他疾病的细胞疗法.
在低pH条件下对调节细胞存活的基因产生了宝贵的知识.

关键词:

通过激活CRISPR来进行激活.细胞疗法细胞疗法细胞活力细胞的活力.组织工程是组织工程.

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