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CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

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CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

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Updated: Feb 18, 2026

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

Combinatorial metabolic engineering using an orthogonal tri-functional CRISPR system.

Jiazhang Lian^1,2, Mohammad HamediRad¹, Sumeng Hu¹

¹Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.

Nature Communications

|November 24, 2017

Summary

This summary is machine-generated.

This study introduces a CRISPR-AID system for yeast metabolic engineering, enabling high-throughput gene manipulation. This combinatorial approach accelerates the optimization of microbial cell factories for improved product yields.

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Area of Science:

Synthetic Biology
Metabolic Engineering
CRISPR Technology

Background:

Microbial cell factory optimization requires complex genetic modifications like gene overexpression, knock-down, and knock-out.
Current methods for these genetic alterations are often sequential and low-throughput, hindering rapid strain development.

Purpose of the Study:

To develop a high-throughput, combinatorial metabolic engineering strategy for yeast.
To enable simultaneous perturbation of multiple gene targets using a novel CRISPR system.

Main Methods:

Development of an orthogonal tri-functional CRISPR system (CRISPR-AID) in Saccharomyces cerevisiae.
CRISPR-AID combines transcriptional activation, transcriptional interference, and gene deletion for modular genetic control.
Application of the system for parallel and high-throughput manipulation of metabolic and regulatory networks.

Main Results:

Demonstrated a 3-fold increase in β-carotene production in a single step using CRISPR-AID.
Achieved a 2.5-fold improvement in endoglucanase display on yeast surface through combinatorial optimization.
Successfully optimized multiple metabolic engineering targets simultaneously.

Conclusions:

The CRISPR-AID system offers a modular, parallel, and high-throughput approach for microbial cell factory engineering.
This strategy significantly accelerates the optimization process for enhanced bioproducts and biocatalyst development.