Over the last years, the discovery of various natural and the development of a row of engineered CRISPR/Cas nucleases have made almost every site of plant genomes accessible for the induction of specific changes. Newly developed tools open up a wide range of possibilities for the induction of genetic variability, from changing a single bp to Mbps, and thus to fine-tune plant performance. Whereas early approaches focused on targeted mutagenesis, recently developed tools enable the induction of precise and predefined genomic modifications. The use of base editors allows the substitution of single nucleotides, whereas the use of prime editors and gene targeting methods enables the induction of larger sequence modifications from a few bases to several kbp. Recently, through CRISPR/Cas-mediated chromosome engineering, it became possible to induce heritable inversions and translocations in the Mbp range. Thus, a novel way of breaking and fixing genetic linkages has come into reach for breeders. In addition, sequence-specific recruitment of various factors involved in transcriptional and post-transcriptional regulation has been shown to provide an additional class of methods for the fine tuning of plant performance. In this review, we provide an overview of the most recent progress in the field of CRISPR/Cas-based tool development for plant genome engineering and try to evaluate the importance of these developments for breeding and biotechnological applications.

中文翻译：

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用于微调植物性能的精密 CRISPR/Cas 工具

在过去的几年里，各种天然基因的发现和一系列工程化 CRISPR/Cas 核酸酶的开发，使得几乎植物基因组的每个位点都可以用于诱导特定的变化。新开发的工具为诱导遗传变异开辟了广泛的可能性，从改变单个 bp 到 Mbps，从而微调植物性能。早期的方法侧重于靶向诱变，而最近开发的工具能够诱导精确和预定义的基因组修饰。碱基编辑器的使用允许替换单个核苷酸，而主要编辑器和基因打靶方法的使用能够诱导从几个碱基到几个 kbp 的更大序列修改。最近，通过 CRISPR/Cas 介导的染色体工程，在 Mbp 范围内诱导可遗传倒位和易位成为可能。因此，一种打破和修复遗传联系的新方法已被育种者所接受。此外，参与转录和转录后调控的各种因子的序列特异性募集已被证明为植物性能的微调提供了另一类方法。在这篇综述中，我们概述了基于 CRISPR/Cas 的植物基因组工程工具开发领域的最新进展，并尝试评估这些进展对育种和生物技术应用的重要性。转录和转录后调控中涉及的各种因子的序列特异性募集已被证明为微调植物性能提供了另一类方法。在这篇综述中，我们概述了基于 CRISPR/Cas 的植物基因组工程工具开发领域的最新进展，并尝试评估这些进展对育种和生物技术应用的重要性。转录和转录后调控中涉及的各种因子的序列特异性募集已被证明为微调植物性能提供了另一类方法。在这篇综述中，我们概述了基于 CRISPR/Cas 的植物基因组工程工具开发领域的最新进展，并尝试评估这些进展对育种和生物技术应用的重要性。