Abstract:
Transgenesis technologies, such as overexpression or RNA interference-mediated suppression, have often been used to alter the activity of target genes. More recently developed targeted genome modification methods using customizable endonucleases allow for the regulation or knockout mutation of target genes without the necessity of integrating recombinant DNA. Such approaches make it possible to create novel alleles of target genes, thereby significantly contributing to crop improvement. Among these technologies, the Cas9 endonuclease-based method is widely applied to several crops, including barley (Hordeum vulgare). In this chapter, we describe an Agrobacterium-based approach to the targeted modification of grain dormancy genes in barley using RNA-guided Cas9 nuclease.
摘要:
转基因技术,如过度表达或RNA干扰介导的抑制,经常被用来改变靶基因的活性。最近开发的使用可定制的核酸内切酶的靶向基因组修饰方法允许靶基因的调节或敲除突变,而不需要整合重组DNA。这样的方法使得产生新的靶基因等位基因成为可能,从而显著促进作物改良。在这些技术中,基于Cas9核酸内切酶的方法广泛应用于几种作物,包括大麦(大麦)。在这一章中,我们描述了一种基于农杆菌的方法,使用RNA指导的Cas9核酸酶靶向修饰大麦中的谷物休眠基因。