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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 and crRNAs

CRISPR and crRNAs

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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...

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.
The recognition sites for Cre recombinase called LoxP...

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Related Experiment Video

Updated: Jul 5, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Programmable RNA base editing with photoactivatable CRISPR-Cas13.

Jeonghye Yu¹, Jongpil Shin¹, Jihwan Yu¹

¹Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.

Nature Communications

|January 22, 2024

Summary

This summary is machine-generated.

Researchers developed a light-inducible CRISPR-Cas13 system (paCas13) for RNA manipulation. This system enables precise control over RNA degradation and base editing in cells and animal models, offering new therapeutic avenues.

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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

Area of Science:

Molecular Biology
Gene Editing Technologies
Biotechnology

Background:

CRISPR-Cas13 systems are versatile tools for RNA manipulation, including interference, imaging, and editing.
Existing CRISPR-Cas13 systems lack precise temporal and spatial control over their activity.
Development of inducible systems is crucial for precise biological research and therapeutic applications.

Purpose of the Study:

To develop a light-inducible CRISPR-Cas13 system for spatiotemporal control of RNA modulation.
To create a light-inducible base editor for reversible RNA editing.
To demonstrate the in vitro and in vivo applicability of these novel RNA-modulating systems.

Main Methods:

Development of a split-Cas13 system (paCas13) fused with Magnet, identifying the optimal N351/C350 split site.
Fusion of ADAR2 with catalytically inactive paCas13 fragments to create a light-inducible base editor (padCas13).
Testing of paCas13 and padCas13 systems for RNA perturbation, base editing (A-to-I, C-to-U), and transcript modulation in mammalian cells and a mouse model.

Main Results:

The paCas13 system demonstrated high inducibility and low background activity for light-induced RNA perturbation.
The padCas13 editor enabled reversible RNA base editing (A-to-I, C-to-U) under light control in vitro.
The padCas13 editor successfully adjusted post-translational modifications and activated target transcripts in vivo in a mouse model.

Conclusions:

A novel light-inducible CRISPR-Cas13 system (paCas13) and a base editor (padCas13) were successfully developed.
These systems offer precise, light-dependent control over RNA degradation and base editing in vitro and in vivo.
The paCas13 system holds broad applicability for RNA manipulation in diverse disease states and physiological processes, advancing research and therapeutic development.