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Updated: Jun 24, 2025

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

Molecular recording using DNA Typewriter.

Hanna Liao^1,2, Junhong Choi^3,4, Jay Shendure^5,6,7,8

¹Department of Genome Sciences, University of Washington, Seattle, WA, USA.

Nature Protocols

|June 6, 2024

Summary

This summary is machine-generated.

The DNA Typewriter system uses prime editing to record cellular history in DNA. This molecular recording method captures temporal relationships for applications in development and cancer evolution research.

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Published on: August 15, 2018

Area of Science:

Genetics
Molecular Biology
Bioengineering

Background:

Molecular recording in genomic DNA offers insights into cellular processes like development and cancer.
Existing genome editing methods struggle to capture temporal relationships of recorded events in mammalian cells.

Purpose of the Study:

To introduce and detail the DNA Typewriter system for high-capacity temporal molecular recording in mammalian cells.
To provide a protocol for implementing DNA Typewriter for recording transfection events and cellular lineage.

Main Methods:

The DNA Typewriter system utilizes prime editing for sequential insertions into an engineered "DNA Tape" locus.
The protocol involves designing DNA Typewriter components, performing recording experiments in HEK293T cells, and analyzing sequencing data.

Main Results:

Demonstrated recording of sequential transfection events using programmed barcode insertions.
Successfully captured lineage information during single-cell expansion and proliferation.
Achieved higher recording capacities for temporal information compared to existing methods.

Conclusions:

DNA Typewriter enables robust temporal molecular recording in mammalian cells.
The system facilitates the study of complex biological processes with high-resolution historical data.
This protocol offers a feasible method for researchers with basic molecular biology expertise.