Whole-organism lineage tracing by combinatorial and cumulative genome editing.
Whole-organism lineage tracing by combinatorial and cumulative genome editing.
复制标题
DOI：
10.1126/science.aaf7907
复制
发表时间：
2016-07-29
期刊：
影响因子：
--
通讯作者：
Shendure J
中科院分区：
文献类型：
--
作者：
McKenna A;Findlay GM;Gagnon JA;Horwitz MS;Schier AF;Shendure J
多细胞系统从单细胞通过不同的谱系发展而来。然而，目前的谱系追踪方法对整个复杂生物的规模很小。在这里，我们使用基因组编辑在多轮细胞分裂中逐步引入和积累DNA条形码中的不同突变。条形码是CRISPR/Cas9靶位点的阵列，标记细胞，并通过细胞之间共享的突变模式阐明谱系关系。在细胞培养和斑马鱼中，我们表明编辑的速率和模式是可调的，并且可以生成数千个谱系信息条形码等位基因。通过从斑马鱼个体中取样数十万个细胞，我们发现成年器官中的大多数细胞来自相对较少的胚胎祖细胞。在未来的分析中，用于谱系追踪的合成靶阵列（GESTALT）的基因组编辑可用于生成多细胞系统中正常发育和疾病的细胞谱系的大规模图谱。
Multicellular systems develop from single cells through distinct lineages. However, current lineage tracing approaches scale poorly to whole, complex organisms. Here we use genome editing to progressively introduce and accumulate diverse mutations in a DNA barcode over multiple rounds of cell division. The barcode, an array of CRISPR/Cas9 target sites, marks cells and enables the elucidation of lineage relationships via the patterns of mutations shared between cells. In cell culture and zebrafish, we show that rates and patterns of editing are tunable, and that thousands of lineage-informative barcode alleles can be generated. By sampling hundreds of thousands of cells from individual zebrafish, we find that most cells in adult organs derive from relatively few embryonic progenitors. In future analyses, genome editing of synthetic target arrays for lineage tracing (GESTALT) can be used to generate large-scale maps of cell lineage in multicellular systems for normal development and disease.