Lineage tracing using a Cas9-deaminase barcoding system targeting endogenous L1 elements

• DOI: 10.1038/s41467-019-09203-z
• Publication Date: 2019-03-15T11:02:32Z
• Authors: Byungjin Hwang; Wookjae Lee; Soo-Young Yum; Yujin Jeon; Namjin Cho; Goo Jang; Duhee Bang
• Keywords: Lineage (genetic); Cell fate determination

Abstract Determining cell lineage and function is critical to understanding human physiology pathology. Although advances in tracing methods provide new insight into fate, defining cellular diversity at the mammalian level remains a challenge. Here, we develop genome editing strategy using cytidine deaminase fused with nickase Cas9 (nCas9) specifically target endogenous interspersed repeat regions cells. The resulting mutation patterns serve as genetic barcode, which induced by targeted mutagenesis single-guide RNA (sgRNA), leveraging substitution events, subsequent read out single primer pair. By analyzing signatures, show accurate reconstruction of both bulk single-cell data. We envision that our barcode system will enable fine-resolution mapping organismal development healthy diseased states.