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Transgenic Plants

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Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
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Related Experiment Video

Updated: Jan 10, 2026

Author Spotlight: Optimized Transformation Protocol for Chlorella vulgaris Using Agrobacterium tumefaciens

Author Spotlight: Optimized Transformation Protocol for Chlorella vulgaris Using Agrobacterium tumefaciens

Published on: October 27, 2023

IMAGE: INTEGRATE-Mediated Agrobacterium Genome Engineering.

Ephraim Aliu^1,2,3, Liang-Chun Chen^1,2,3, Keunsub Lee^1,2

¹Department of Agronomy, Iowa State University, Ames, IA, United States.

Frontiers in Microbiology

|November 24, 2025

Summary

This summary is machine-generated.

Researchers optimized the CRISPR RNA-guided transposase system, INTEGRATE, for precise Agrobacterium genome engineering. This standardized protocol facilitates Agrobacterium strain development for plant biotechnology applications.

Keywords:

Agrobacterium rhizogenes Agrobacterium spp.Agrobacterium tumefaciens CRISPR integrase Cre-lox disarming large DNA fragment deletion targeted DNA insertion

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Last Updated: Jan 10, 2026

Author Spotlight: Optimized Transformation Protocol for Chlorella vulgaris Using Agrobacterium tumefaciens

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Published on: October 27, 2023

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Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)

Published on: March 16, 2012

Area of Science:

Microbiology
Molecular Biology
Biotechnology

Background:

Precise engineering of Agrobacterium strains is vital for advancing plant biotechnology.
The CRISPR RNA-guided transposase system, INTEGRATE, offers an efficient method for genetic modification in Agrobacterium.
Current applications of INTEGRATE in Agrobacterium strain engineering require further exploration and standardization.

Purpose of the Study:

To present a standardized and optimized workflow for utilizing the INTEGRATE system in Agrobacterium.
To address and overcome common challenges associated with INTEGRATE implementation, including crRNA design, transformation efficiency, and vector eviction.
To expand the genetic engineering toolkit for Agrobacterium to support functional genomics and strain development.

Main Methods:

Development of a standardized protocol for CRISPR-RNA guided transposase (INTEGRATE) system application in Agrobacterium.
Optimization of target site selection, crRNA spacer cloning, and Agrobacterium transformation procedures.
Implementation of screening methods for targeted insertion, Cre/loxP-mediated deletion, and vector removal.

Main Results:

A robust workflow for targeted genome modifications in Agrobacterium using the INTEGRATE system was established.
The protocol successfully demonstrated the domestication of Agrobacterium rhizogenes K599 by deleting the T-DNA region and creating auxotrophy.
The developed methods enhance efficiency and reproducibility of Agrobacterium genome engineering.

Conclusions:

The optimized INTEGRATE protocol provides a valuable resource for researchers performing Agrobacterium genome engineering.
This standardized workflow facilitates the development of improved Agrobacterium strains for plant transformation applications.
The study promotes broader adoption of advanced genetic engineering tools in Agrobacterium, driving innovation in plant biotechnology.