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Related Experiment Video

Updated: Jan 26, 2026

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Published on: November 1, 2019

Targeted Gene Activation Using RNA-Guided Nucleases.

Alexander Brown¹, Wendy S Woods¹, Pablo Perez-Pinera²

¹Department of Bioengineering, University of Illinois at Urbana-Champaign, 1270 Digital Computer Laboratory, MC-278, 1304 West Springfield Avenue, Urbana, IL, 61801-2910, USA.

Methods in Molecular Biology (Clifton, N.J.)

|September 25, 2016

Summary

This summary is machine-generated.

CRISPR-Cas9 transcription factors enable precise gene activation. This chapter details methods for designing, constructing, and analyzing these powerful synthetic gene regulators in mammalian cells.

Keywords:

CRISPR-Cas9 Gene activation Gene expression Genome engineering RNA-guided nucleases Synthetic biology Transcription

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Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells

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Published on: August 23, 2014

Area of Science:

Molecular Biology
Genome Engineering
Synthetic Biology

Background:

The CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) system has revolutionized genome engineering.
Catalytically inactive Cas9 fused with transcriptional activators creates customizable synthetic transcription factors.
These factors allow robust, specific, and tunable gene expression activation.

Purpose of the Study:

To describe experimental design considerations for CRISPR-Cas transcription factors.
To outline methods for plasmid construction and gene delivery.
To detail procedures for analyzing targeted gene activation in mammalian cells.

Main Methods:

Utilizing CRISPR-Cas9 technology for targeted gene activation.
Designing and constructing plasmids with catalytically inactive Cas9 fused to activator domains.
Implementing gene delivery protocols for mammalian cell lines.
Analyzing gene expression changes to confirm targeted activation.

Main Results:

Demonstration of customizable synthetic transcription factors for gene activation.
Establishment of robust, specific, and tunable gene expression control.
Successful application in a variety of mammalian tissues and cells.

Conclusions:

CRISPR-Cas transcription factors offer a powerful and versatile tool for genome engineering.
The described methods facilitate the design and analysis of targeted gene activation strategies.
This approach enables precise control over gene expression in mammalian systems.