Department of Molecular Biotechnology and Bioinformatics, Università degli Studi di Milano, Milan, Italy.
Division of Neuroscience, Department of Pharmacology & Toxicology, Vita-Salute San Raffaele University and Hospital, Milan, Italy.
Mol Biotechnol. 2023 Feb;65(2):196-205. doi: 10.1007/s12033-022-00538-5. Epub 2022 Aug 8.
CRISPR-Cas systems play a critical role in the prokaryotic adaptive immunity against mobile genetic elements, such as phages and foreign plasmids. In the last decade, Cas9 has been established as a powerful and versatile gene editing tool. In its wake, the novel RNA-guided endonuclease system CRISPR-Cas12a is transforming biological research due to its unique properties, such as its high specificity or its ability to target T-rich motifs, to induce staggered double-strand breaks and to process RNA arrays. Meanwhile, there is an increasing need for efficient and safe gene activation, repression or editing in pluricellular organisms for crop improvement, gene therapy, research model development, and other goals. In this article, we review CRISPR-Cas12a applications in pluricellular organisms and discuss how the challenges characteristic of these complex models, such as vectorization or temperature variations in ectothermic species, can be overcome.
CRISPR-Cas 系统在原核生物对抗移动遗传元件(如噬菌体和外源质粒)的适应性免疫中起着关键作用。在过去的十年中,Cas9 已被确立为一种强大而多功能的基因编辑工具。继 Cas9 之后,新型的 RNA 指导内切酶 Cas12a 因其独特的特性,如高特异性或靶向富含 T 的基序的能力、诱导交错双链断裂以及处理 RNA 阵列的能力,正在改变生物研究。同时,对于农作物改良、基因治疗、研究模型开发和其他目标,在多细胞生物中高效、安全地激活、抑制或编辑基因的需求也在不断增加。在本文中,我们回顾了 Cas12a 在多细胞生物中的应用,并讨论了如何克服这些复杂模型的特征性挑战,如载体化或变温动物物种中的温度变化。