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


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

SUBMITTER: Hwang B 

PROVIDER: S-EPMC6420643 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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

Hwang Byungjin B   Lee Wookjae W   Yum Soo-Young SY   Jeon Yujin Y   Cho Namjin N   Jang Goo G   Bang Duhee D  

Nature communications 20190315 1


Determining cell lineage and function is critical to understanding human physiology and pathology. Although advances in lineage tracing methods provide new insight into cell fate, defining cellular diversity at the mammalian level remains a challenge. Here, we develop a genome editing strategy using a cytidine deaminase fused with nickase Cas9 (nCas9) to specifically target endogenous interspersed repeat regions in mammalian cells. The resulting mutation patterns serve as a genetic barcode, whic  ...[more]

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