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

Byungjin Hwang, Wookjae Lee, Soo Young Yum, Yujin Jeon, Namjin Cho, Goo Jang, Duhee Bang

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Article number1234
JournalNature communications
Volume10
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

Fingerprint

Guide RNA
Long Interspersed Nucleotide Elements
Cytidine Deaminase
Mutagenesis
Deoxyribonuclease I
Physiology
Pathology
tracing
Substitution reactions
Genes
Cells
Cell Lineage
cells
mutations
Mutation
mutagenesis
editing
primers
physiology
genome

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Hwang, Byungjin ; Lee, Wookjae ; Yum, Soo Young ; Jeon, Yujin ; Cho, Namjin ; Jang, Goo ; Bang, Duhee. / Lineage tracing using a Cas9-deaminase barcoding system targeting endogenous L1 elements. In: Nature communications. 2019 ; Vol. 10, No. 1.
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Lineage tracing using a Cas9-deaminase barcoding system targeting endogenous L1 elements. / Hwang, Byungjin; Lee, Wookjae; Yum, Soo Young; Jeon, Yujin; Cho, Namjin; Jang, Goo; Bang, Duhee.

In: Nature communications, Vol. 10, No. 1, 1234, 01.12.2019.

Research output: Contribution to journalArticle

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