利用 CRISPR/Cas9 在多倍体中进行基因组编辑。
Genome Editing by CRISPR/Cas9 in Polyploids.
机构信息
Departamento de Mejora Genética y Biotecnología, Instituto de Hortofruticultura Subtropical y Mediterránea (IHSM), Universidad de Málaga - Consejo Superior de Investigaciones Científicas, Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, UMA, Málaga, Spain.
出版信息
Methods Mol Biol. 2023;2545:459-473. doi: 10.1007/978-1-0716-2561-3_24.
CRISPR/Cas system has been widely used for genome editing in the past few years. Even though it has been performed in many polyploid species to date, its efficient accomplishment in these organisms is still a challenge. The presence of multiple homoeologous genes as targets for their editing requires more rigorous work and specific needs to assess successful genome editing. Here, we describe a general stepwise protocol to select target sites, design sgRNAs, indicate vector requirements, and screen CRISPR/Cas9-mediated genome editing in polyploid species.
CRISPR/Cas 系统在过去几年中被广泛用于基因组编辑。尽管迄今为止已经在许多多倍体物种中进行了这项工作，但在这些生物体中有效地完成它仍然是一个挑战。作为编辑目标的多个同源基因的存在需要更严格的工作和特定的需求来评估成功的基因组编辑。在这里，我们描述了一个通用的逐步方案，用于选择靶位点、设计 sgRNA、指示载体要求，并筛选多倍体物种中的 CRISPR/Cas9 介导的基因组编辑。
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