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Related Experiment Video

Updated: Oct 3, 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

CRISPR-Act3.0-Based Highly Efficient Multiplexed Gene Activation in Plants.

Changtian Pan¹, Yiping Qi^1,2

¹Department of Plant Science and Landscape Architecture, University of Maryland, College Park, Maryland.

Current Protocols

|February 14, 2022

Summary

This summary is machine-generated.

This study presents a simple CRISPR-Act3.0 protocol for multiplexed gene activation in rice. This CRISPR/Cas system allows targeted gene activation without DNA breaks, aiding research in gene networks and metabolic pathways.

Keywords:

CRISPR-Act3.0 dCas9 multiplexed gene activation rice protoplast system sgRNA design tRNA

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Last Updated: Oct 3, 2025

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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

Area of Science:

Molecular Biology
Plant Biotechnology
Genome Engineering

Background:

CRISPR/Cas systems are powerful tools for genome editing and transcriptional regulation.
Catalytically inactive Cas variants (dCas) fused with activators enable targeted gene activation without DNA double-strand breaks.
Multiplexed gene activation holds potential for understanding gene regulatory networks and metabolic engineering.

Purpose of the Study:

To describe a simple protocol for constructing a dCas9-mediated multiplexed gene activation system.
To evaluate the CRISPR-Act3.0 system for its efficiency in rice protoplasts.
To provide a method for simultaneous activation of multiple genes.

Main Methods:

Design of single-guide RNA (sgRNA) for target gene specificity.
Construction of CRISPR-Act3.0 vectors for multiplexed gene activation.
Testing vector efficiency in rice protoplasts.

Main Results:

Successful construction of dCas9-mediated multiplexed gene activation vectors.
Demonstration of CRISPR-Act3.0 system functionality in rice protoplasts.
Validation of the protocol for activating multiple genes simultaneously.

Conclusions:

The developed CRISPR-Act3.0 protocol offers a straightforward method for multiplexed gene activation in rice.
This system provides a valuable tool for dissecting gene regulatory networks and metabolic pathways.
The technology facilitates advancements in plant functional genomics and breeding.