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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: Mar 17, 2026

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Tissue-specific gene targeting using CRISPR/Cas9.

J Ablain¹, L I Zon¹

¹Howard Hughes Medical Institute and Harvard Medical School, Boston, MA, United States.

Methods in Cell Biology

|July 23, 2016

Summary

This summary is machine-generated.

CRISPR/Cas9 technology enables efficient gene inactivation in zebrafish for studying gene function. This chapter details a method for tissue-specific gene targeting, overcoming limitations of whole-embryo silencing.

Keywords:

CRISPR/Cas9 technology Gene knockout Morpholino Tissue specificity gRNAs

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Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Published on: February 28, 2025

Area of Science:

Developmental Biology
Genetics
Molecular Biology

Background:

Zebrafish are crucial models for genetic screens in organogenesis and development.
Reverse genetics in zebrafish historically lacked efficient knockout technologies, hindering adult studies.
CRISPR/Cas9 has advanced loss-of-function studies but lacks spatial control for tissue-specific inactivation.

Purpose of the Study:

To present a method for tissue-specific gene inactivation in zebrafish using CRISPR/Cas9.
To highlight the advantages of spatial gene targeting over whole-embryo silencing.
To discuss applications and limitations of this advanced technique.

Main Methods:

Utilizing CRISPR/Cas9 genome editing for targeted gene modification.
Developing a strategy for achieving tissue-specific gene inactivation in zebrafish.
Phenotypic analysis of gene silencing in specific tissues.

Main Results:

Demonstration of a method for precise, tissue-specific gene targeting in zebrafish.
Overcoming challenges associated with spatial control in gene inactivation studies.
Facilitating advanced loss-of-function studies in adult zebrafish.

Conclusions:

CRISPR/Cas9-mediated tissue-specific gene inactivation is a powerful tool for zebrafish research.
This method addresses limitations of previous techniques, enabling detailed functional studies.
Future developments promise enhanced spatial and temporal control for gene targeting.