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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...

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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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CRISPR and crRNAs

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Related Experiment Video

Updated: Aug 28, 2025

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

Transposons and CRISPR: Rewiring Gene Editing.

Francisco Tenjo-Castaño¹, Guillermo Montoya¹, Arturo Carabias¹

¹Structural Molecular Biology Group, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3-B, Copenhagen 2200, Denmark.

|September 21, 2022

Summary

New transposon-associated RNA-guided systems offer innovative CRISPR-Cas gene editing solutions. These OMEGA nucleases and RNA-guided transposons show promise for overcoming limitations in current gene editing technologies.

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Last Updated: Aug 28, 2025

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

CRISPR-Cas gene editing is revolutionary but limited by off-target effects and reliance on specific DNA repair pathways.
Current gene editing methods struggle with efficiency in non-dividing cells and insertion of large DNA payloads.

Purpose of the Study:

To explore novel transposon-associated RNA-guided mechanisms as potential gene editing solutions.
To assess the potential of OMEGA nucleases and RNA-guided transposons in overcoming current gene editing challenges.

Main Methods:

Analysis of IS200/IS605 family transposon-associated endonucleases (OMEGA nucleases) and their comparison to CRISPR-Cas systems.
Investigation of RNA-guided activity in Tn7-like transposons for targeted integration.
Evaluation of gene editing capabilities in human cells.

Main Results:

OMEGA nucleases, derived from transposons, demonstrate gene editing in human cells and offer potential for compact RNA-guided platforms.
RNA-guided transposons, including Tn7-like elements, utilize sophisticated mechanisms for target site selection and integration.
These systems present alternative scaffolds to current CRISPR-Cas technologies.

Conclusions:

Transposon-associated RNA-guided systems represent a promising frontier in gene editing.
OMEGA nucleases and RNA-guided transposons may circumvent limitations of traditional CRISPR-Cas approaches, enhancing gene editing efficiency and versatility.