In the field of tissue engineering, autologous cell sources are ideal to prevent adverse immune responses; however, stable and reliable cell sources are limited. To acquire more reliable cell sources, the harvesting and differentiation of stem cells from patients is becoming more and more common. To this end, the need to control the fate of these stem cells before transplantation for therapeutic purposes is urgent. Since transcription factors orchestrate all of the gene activities inside of a cell, researchers have developed engineered and synthetic transcription factors to precisely control the fate of stem cells allowing for safer and more effective cell sources. Engineered transcription factors, mutant fusion proteins of naturally occurring proteins, comprise the three main domains of natural transcription factors including DNA binding domains, transcriptional activation domains, and a linker domain. Several key advancements of engineered zinc finger proteins, transcriptional activator-like effectors, and deficient cas9 proteins have revolutionized the field of engineered transcription factors allowing for precise control of gene regulation. Synthetic transcription factors are chemically made transcription factor mimics that use small molecule based moieties to replicate the main functions of natural transcription factors. These include hairpin polyamides, triple helix forming oligonucleotides, and nanoparticle-based methods. Synthetic transcription factors allow for non-viral delivery and greater spatiotemporal control of gene expression. The developments in engineered and synthetic transcription factors have lowered the risk of tumorigenicity and improved differentiation capability of stem cells, as well as facilitated many key discoveries in the fields of cancer and stem cell biology, thus providing a stepping stone to advance regenerative medicine in the clinic for cell replacement therapies.

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干细胞治疗学的高级基因操纵方法

在组织工程领域，自体细胞来源是防止不良免疫反应的理想选择。但是，稳定和可靠的细胞来源是有限的。为了获得更可靠的细胞来源，来自患者的干细胞的收获和分化变得越来越普遍。为此，迫切需要在移植之前控制这些干细胞的命运以达到治疗目的。由于转录因子能够协调细胞内所有基因的活动，因此研究人员开发了工程化和合成的转录因子来精确控制干细胞的命运，从而获得更安全，更有效的细胞来源。工程转录因子是天然存在的蛋白质的突变融合蛋白，包含天然转录因子的三个主要域，包括DNA结合域，转录激活结构域和接头结构域。工程锌指蛋白，类似转录激活因子的效应子和cas9蛋白不足的几个关键进展彻底改变了工程转录因子领域，从而可以精确控制基因调控。合成转录因子是化学合成的转录因子模拟物，它使用基于小分子的部分复制天然转录因子的主要功能。这些包括发夹状聚酰胺，三螺旋形成寡核苷酸和基于纳米粒子的方法。合成转录因子允许非病毒传递和更大的时空控制基因表达。