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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...

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Related Experiment Video

Updated: May 24, 2025

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

Developing safe and efficient CGBE editor based on Cas-embedding strategy.

Tian Lin^1,2,3, Xin Wang⁴, Yu Zhang^4,5

¹Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou, Jiangsu, 221004, China.

Synthetic and Systems Biotechnology

|March 3, 2025

Summary

This summary is machine-generated.

We developed novel Cas-embedding C-to-G base editors (CE-CGBE) for precise DNA editing. The HF-CGBE variant shows high efficiency and purity with minimal off-target effects, offering therapeutic potential.

Keywords:

CGBE Cas-embedding Off-target eA3A

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Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Published on: February 28, 2021

Area of Science:

Molecular Biology
Gene Editing Technologies
Biochemistry

Background:

Cytosine-to-guanine base editors (CGBE) precisely convert cytosines to guanines.
Conventional CGBE systems face challenges due to non-specific DNA binding of cytidine deaminases, causing off-target effects.
The Cas-embedding strategy integrates functional proteins into Cas9 to enhance specificity and reduce off-target mutations.

Purpose of the Study:

To pioneer the Cas-embedding strategy for CGBE systems.
To engineer novel CGBE editors, termed CE-CGBE, with improved safety and efficacy.
To evaluate the performance of engineered CE-CGBE editors, particularly the HF-CGBE variant.

Main Methods:

Application of the Cas-embedding strategy to CGBE systems.
Engineering of novel CE-CGBE editors incorporating eA3A, RBMX, and Udgx.
Assessment of editing efficiency, purity, indel formation, and off-target effects (DNA and RNA) for HF-CGBE.

Main Results:

Engineered CE-CGBE editors demonstrated high editing efficiency and purity.
The HF-CGBE variant, incorporating eA3A, RBMX, and Udgx, exhibited superior performance.
HF-CGBE showed no significant difference in off-target effects compared to negative controls for both DNA and RNA.

Conclusions:

Novel HF-CGBE editors significantly expand the base editing toolbox.
The developed CE-CGBE systems offer a promising therapeutic avenue for pathogenic mutations.
Cas-embedding strategy effectively mitigates off-target effects in CGBE systems.