Abstract:
Duplication is a foundation of molecular evolution and a driver of genomic and complex diseases. Here, we develop a genome editing tool named Amplification Editing (AE) that enables programmable DNA duplication with precision at chromosomal scale. AE can duplicate human genomes ranging from 20 bp to 100 Mb, a size comparable to human chromosomes. AE exhibits activity across various cell types, encompassing diploid, haploid, and primary cells. AE exhibited up to 73.0% efficiency for 1 Mb and 3.4% for 100 Mb duplications, respectively. Whole-genome sequencing and deep sequencing of the junctions of edited sequences confirm the precision of duplication. AE can create chromosomal microduplications within disease-relevant regions in embryonic stem cells, indicating its potential for generating cellular and animal models. AE is a precise and efficient tool for chromosomal engineering and DNA duplication, broadening the landscape of precision genome editing from an individual genetic locus to the chromosomal scale.
摘要:
复制是分子进化的基础,也是基因组和复杂疾病的驱动因素。这里,我们开发了一种名为扩增编辑(AE)的基因组编辑工具,该工具可以在染色体尺度上精确地进行可编程的DNA复制。AE可以复制20bp至100Mb的人类基因组,与人类染色体相当的大小。AE在各种细胞类型中表现出活性,包括二倍体,单倍体,和原代细胞。1Mb的AE效率高达73.0%,100Mb的AE效率高达3.4%,分别。编辑序列的连接的全基因组测序和深度测序证实了复制的精确性。AE可以在胚胎干细胞的疾病相关区域内产生染色体微复制,表明其产生细胞和动物模型的潜力。AE是一种精确有效的染色体工程和DNA复制工具,将精确基因组编辑的前景从个体遗传基因位点扩展到染色体尺度。