Abstract：
Transcriptional activation, based on Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) and known as CRISPR activation (CRISPRa), is a specific and safe tool to upregulate endogenous genes. Therefore, CRISPRa is valuable not only for analysis of molecular mechanisms of cellular events, but also for treatment of various diseases. Regulating autophagy has been proposed to enhance effects of some therapies. In this study, we upregulated genes for phosphoinositide phosphatases, SACM1L, PIP4P1, and PIP4P2, using CRISPRa, and their effects on autophagy were examined. Our results suggested that TMEM55A/PIP4P2, a phosphatidylinositol-4,5-bisphosphate 4-phosphatase, positively regulates basal autophagy in 293A cells. Furthermore, it was also suggested that SAC1, a phosphatidylinositol 4-phosphatase, negatively regulates basal autophagic degradation.
摘要：
转录激活,基于聚集的规则间隔短回文重复序列（CRISPR）-CRISPR相关蛋白9（Cas9），称为CRISPR激活（CRISPRa），是上调内源基因的特异性和安全工具。因此,CRISPRa不仅对分析细胞事件的分子机制有价值，还用于治疗各种疾病。已经提出调节自噬以增强一些疗法的效果。在这项研究中,我们上调了磷酸肌醇磷酸酶的基因，SACM1L,PIP4P1和PIP4P2，使用CRISPRa，并检查了它们对自噬的影响。我们的结果表明TMEM55A/PIP4P2，一种磷脂酰肌醇-4,5-二磷酸4-磷酸酶，在293A细胞中正向调节基础自噬。此外，还建议SAC1，一种磷脂酰肌醇4-磷酸酶，负调节基础自噬降解。
