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Lineage tracing and analog recording in mammalian cells by single-site DNA writing
- Source :
- Nature chemical biology, vol 17, iss 6, Nat Chem Biol
- Publication Year :
- 2021
- Publisher :
- eScholarship, University of California, 2021.
-
Abstract
- Studying cellular and developmental processes in complex multicellular organisms can require the non-destructive observation of thousands to billions of cells deep within an animal. DNA recorders address the staggering difficulty of this task by converting transient cellular experiences into mutations at defined genomic sites that can be sequenced later in high throughput. However, existing recorders act primarily by erasing DNA. This is problematic because, in the limit of progressive erasure, no record remains. We present a DNA recorder called CHYRON (Cell History Recording by Ordered Insertion) that acts primarily by writing new DNA through the repeated insertion of random nucleotides at a single locus in temporal order. To achieve in vivo DNA writing, CHYRON combines Cas9, a homing guide RNA and the template-independent DNA polymerase terminal deoxynucleotidyl transferase. We successfully applied CHYRON as an evolving lineage tracer and as a recorder of user-selected cellular stimuli.
- Subjects :
- Biochemistry & Molecular Biology
Lineage (genetic)
DNA polymerase
Cells
1.1 Normal biological development and functioning
Computational biology
DNA-Directed DNA Polymerase
Article
03 medical and health sciences
chemistry.chemical_compound
Medicinal and Biomolecular Chemistry
Underpinning research
Insertional
Genetics
Humans
A-DNA
Cell Lineage
Guide RNA
Kinetoplastida
Molecular Biology
Cells, Cultured
030304 developmental biology
0303 health sciences
Cultured
biology
Cas9
Nucleotides
030302 biochemistry & molecular biology
High-Throughput Nucleotide Sequencing
Cell Biology
DNA
Multicellular organism
Mutagenesis, Insertional
HEK293 Cells
Terminal deoxynucleotidyl transferase
chemistry
Mutagenesis
Mutation
biology.protein
RNA
RNA Editing
Generic health relevance
Biochemistry and Cell Biology
CRISPR-Cas Systems
Guide
RNA, Guide, Kinetoplastida
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Nature chemical biology, vol 17, iss 6, Nat Chem Biol
- Accession number :
- edsair.doi.dedup.....be7ba44077e89c8b13b96bbcf17a64cf