Targeted engineering of plant centromeres

工程小染色体是分析植物染色体功能、生物技术和植物育种的理想工具。在初步研究中，我们开发了一种通过将重组着丝粒蛋白靶向于非着丝粒位置的串联重复序列阵列来重新生成着丝粒的方法，并建立了一种在植物中通过端粒播种进行非靶向染色体切断的方法。现在，我们的目标是结合这两种方法来产生工程化的小染色体。为此，拟在拟南芥中应用靶向新生着丝粒和靶向染色体截断作为实验系统。此外，着丝点组装因子和表观遗传设置是有效的新生着丝点形成所必需的，将被破译。将测定新生着丝粒的有丝分裂和减数分裂活性。

Engineered minichromosomes are highly desirable tools for analysis of plant chromosome function, biotechnology and plant breeding purposes. In preliminary studies, we have developed a method for the de novo generation of centromeres via the targeting of recombinant kinetochore proteins to tandem repeat arrays at non-centromeric positions and established a method for non-targeted chromosome truncation via telomere seeding in plants. Now, we aim to combine both methods for the generation of engineered minichromosomes. For this, targeted de novo centromere and targeted chromosome truncation will be applied as an experimental system in A. thaliana. Further, kinetochore assembly factors and the epigenetic setup which are required for efficient de novo centromere formation will be deciphered. The mitotic and meiotic activity of de novo centromeres will be determined.