Sprink Thorben, Metje Janina, Hartung Frank
Julius Kühn Institut, Institute for Biosafety in Plant Biotechnology, Erwin Baur-Str. 27, 06484 Quedlinburg, Germany.
Max Plank Institute for Biophysical Chemistry, Research Group Autophagy, Am Fassberg 11, 37077 Göttingen, Germany.
Curr Opin Biotechnol. 2015 Apr;32:47-53. doi: 10.1016/j.copbio.2014.11.010. Epub 2014 Nov 25.
Genome editing technologies using sequence specific nucleases (SSNs) became a tremendously powerful and precise tool for reverse genetic approaches and applied biology. Transcription activator-like effector nucleases (TALENs) in particular, consisting of a free designable DNA binding domain and a nuclease, have been exploited today by a huge number of approaches in many different organisms. The convenience of designing the DNA binding domain and straightforward protocols for their assembly, as well as the broad number of applications in different scientific fields made it Natures method of the year 2011. TALENs act as molecular scissors by introducing double strand breaks (DSBs) to the DNA at a given location. The DSBs are subsequently repaired by the cell itself using different repair pathways such as non-homologous end joining (NHEJ) or homologous recombination (HR). These mechanisms can lead to deletions, insertions, replacements or larger chromosomal rearrangements. By offering a template DNA it is possible to channel the repair in direction of HR. In this article we review the recent findings in the field of SSN approaches with emphasis on plants.
使用序列特异性核酸酶（SSN）的基因组编辑技术已成为反向遗传学方法和应用生物学中极为强大且精确的工具。特别是转录激活样效应物核酸酶（TALEN），它由一个可自由设计的DNA结合结构域和一个核酸酶组成，如今已在许多不同生物体中通过大量方法得到应用。其DNA结合结构域设计的便利性、组装的简单流程以及在不同科学领域的广泛应用，使其成为2011年的《自然》年度方法。TALEN通过在特定位置的DNA上引入双链断裂（DSB）来充当分子剪刀。随后，细胞自身利用不同的修复途径，如非同源末端连接（NHEJ）或同源重组（HR）来修复DSB。这些机制可导致缺失、插入、替换或更大规模的染色体重排。通过提供模板DNA，有可能引导修复朝着HR的方向进行。在本文中，我们重点回顾了植物领域中SSN方法的最新研究成果。
