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Related Experiment Video

Updated: Jul 2, 2025

High-throughput CRISPR Vector Construction and Characterization of DNA Modifications by Generation of Tomato Hairy Roots

High-throughput CRISPR Vector Construction and Characterization of DNA Modifications by Generation of Tomato Hairy Roots

Published on: April 30, 2016

A CRISPR interference system for engineering biological nitrogen fixation.

Steven J Russell¹, Amanda K Garcia¹, Betül Kaçar¹

¹Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin, USA.

|February 20, 2024

Summary

This summary is machine-generated.

Researchers developed a CRISPR interference (CRISPRi) system for gene silencing in Azotobacter vinelandii, a key bacterium for biological nitrogen fixation. This bioengineering tool aids in understanding and manipulating nitrogen fixation for climate change solutions.

Keywords:

Azotobacter vinelandii CRISPRi gene silencing nitrogen fixation nitrogenase

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Last Updated: Jul 2, 2025

High-throughput CRISPR Vector Construction and Characterization of DNA Modifications by Generation of Tomato Hairy Roots

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Published on: April 30, 2016

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira

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Published on: August 14, 2021

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis

Published on: June 9, 2020

Area of Science:

Microbiology
Bioengineering
Genetics

Background:

Biological nitrogen fixation is crucial for converting atmospheric nitrogen into bioavailable ammonia, essential for life.
Industrial nitrogen fertilizer production is energy-intensive and environmentally damaging.
Genetic tools for engineering nitrogen-fixing microbes like Azotobacter vinelandii are limited.

Purpose of the Study:

To establish the first CRISPR interference (CRISPRi) system for targeted gene silencing in Azotobacter vinelandii.
To enable advanced genetic manipulation for bioengineering microbial nitrogen fixation.

Main Methods:

Developed a CRISPRi system integrating genomically via site-specific transposon insertion in A. vinelandii.
Demonstrated CRISPRi-mediated repression of an essential nitrogen fixation gene.
Confirmed expression of nitrogenase genes from the transposon insertion site.

Main Results:

Achieved approximately 60% transcriptional repression of a key nitrogen fixation gene using the CRISPRi system.
Validated the functionality of the transposon insertion site for expressing engineered nitrogen fixation genes.
Successfully established a novel gene silencing tool for A. vinelandii.

Conclusions:

The developed CRISPRi system is a significant advancement for studying and engineering nitrogen fixation in A. vinelandii.
This tool facilitates combinatorial studies of gene expression and bioengineering efforts.
It addresses a critical need for genetic tools in microbial nitrogen fixation research, supporting climate change solutions.