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Bacterial Transformation

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In 1928, bacteriologist Frederick Griffith worked on a vaccine for pneumonia, which is caused by Streptococcus pneumoniae bacteria. Griffith studied two pneumonia strains in mice: one pathogenic and one non-pathogenic. Only the pathogenic strain killed host mice.Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...

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Updated: Jul 3, 2026

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Published on: July 7, 2020

High-throughput DNA engineering by mating bacteria.

Takeshi Matsui¹, Po-Hsiang Hung², Han Mei¹

¹BacStitch DNA, Inc., Los Altos, CA, USA; SLAC National Accelerator Laboratory, Menlo Park, CA, USA.

|July 1, 2026

Summary

This summary is machine-generated.

SCRIVENER is a new DNA assembly platform that uses bacterial conjugation and recombination for efficient in vivo DNA engineering. This method simplifies, reduces costs, and increases throughput for creating large DNA constructs.

Keywords:

DNA assembly DNA engineering DNA reuse DNA synthesis composability conjugation genetic tools high-throughput in vivo plasmid engineering

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Area of Science:

Synthetic Biology
Molecular Biology
Biotechnology

Background:

DNA assembly is crucial for synthetic biology and genetic engineering.
Existing methods for DNA assembly can be complex, costly, and time-consuming.
There is a need for streamlined and scalable DNA engineering platforms.

Purpose of the Study:

To introduce SCRIVENER, a novel in vivo DNA assembly platform.
To demonstrate SCRIVENER's ability to streamline and scale DNA engineering.
To provide a simpler, cheaper, and higher-throughput alternative to current DNA assembly methods.

Main Methods:

SCRIVENER utilizes bacterial conjugation, in vivo DNA cutting, and homologous recombination.
DNA blocks are stitched together by mating E. coli in large arrays or pools.
The platform enables assembly without requiring DNA to be moved in and out of cells.

Main Results:

Over 5,000 DNA assemblies were performed with 2 to 19 blocks (240 bp-12 kb).
Constructs up to 81 kb were assembled with high fidelity.
Error rates, primarily deletions between long repeats, were minimized through high-replication assembly and sequence verification.

Conclusions:

SCRIVENER is a powerful tool for accelerating DNA design-build-test-learn cycles.
The platform facilitates combinatorial library construction and DNA block reuse without PCR.
SCRIVENER offers a significant advancement in in vivo DNA engineering capabilities.