Université de Strasbourg, CNRS, INSERM, CELPHEDIA, PHENOMIN, Institut Clinique de la Souris, Illkirch, France.
Methods Mol Biol. 2023;2631:277-297. doi: 10.1007/978-1-0716-2990-1_12.
CRISPR/Cas9 technology is a versatile tool for engineering biology that has dramatically transformed our ability to manipulate genomes. In this protocol, we use its capacity to generate two double-strand breaks simultaneously, at precise positions in the genome, to generate mouse or rat lines with deletion, inversion, and duplication of a specific genomic segment. The technic is called CRISMERE for CRISpr-MEdiated REarrangement. This protocol describes the different steps to generate and validate the different chromosomal rearrangements that can be obtained with the technology. These new genetic configurations can be useful to model rare diseases with copy number variation, understand the genomic organization, or provide genetic tools (like balancer chromosome) to keep lethal mutations.
CRISPR/Cas9 技术是一种用于工程生物学的多功能工具，极大地改变了我们操纵基因组的能力。在本方案中，我们利用其同时在基因组的精确位置产生两个双链断裂的能力，生成小鼠或大鼠的缺失、倒位和特定基因组片段的重复系。该技术称为 CRISPR 介导的重排（CRISMERE）。本方案描述了生成和验证可通过该技术获得的不同染色体重排的不同步骤。这些新的遗传构型可用于模拟具有拷贝数变异的罕见疾病，了解基因组组织，或提供遗传工具（如平衡染色体）来保持致死突变。
