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Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

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

CRISPR/Cas9 Genome Editing

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 18, 2026

Indel Detection following CRISPR/Cas9 Mutagenesis using High-resolution Melt Analysis in the Mosquito Aedes aegypti

Indel Detection following CRISPR/Cas9 Mutagenesis using High-resolution Melt Analysis in the Mosquito Aedes aegypti

Published on: September 10, 2021

CRISP-ID: decoding CRISPR mediated indels by Sanger sequencing.

Jonas Dehairs¹, Ali Talebi¹, Yacine Cherifi²

¹Laboratory of Lipid Metabolism and Cancer, Department of Oncology, KU Leuven, 3000 Leuven, Belgium.

Scientific Reports

|July 2, 2016

Summary

This summary is machine-generated.

Next-generation gene editing tools enable genome engineering, but screening clones with insertions/deletions (indels) is difficult. CRISP-ID is a web tool that genotypes up to three alleles from Sanger sequencing, speeding up clone characterization.

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

Area of Science:

Molecular Biology
Genomics
Bioinformatics

Background:

Next-generation gene editing technologies have advanced genome engineering, enabling gene knockout model creation and functional gene analysis.
Screening gene-edited clones is challenging due to the frequent presence of multiple insertion/deletion (indel) mutations simultaneously.
Efficiently characterizing these indels is crucial for advancing genetic research and therapeutic development.

Purpose of the Study:

To present CRISP-ID, a novel web application designed for the robust genotyping of gene-edited clones.
To provide a user-friendly platform for identifying and characterizing various indels within sequencing data.
To accelerate the process of analyzing gene editing outcomes.

Main Methods:

Development of a unique algorithm for analyzing Sanger sequencing traces.
Implementation of the algorithm within a web application named CRISP-ID.
Capability to genotype up to three distinct alleles from a single sequencing run.

Main Results:

CRISP-ID accurately identifies and differentiates multiple indels within a single Sanger sequencing trace.
The application significantly speeds up the characterization of gene-edited clones compared to traditional methods.
Provides a robust and accessible solution for genotyping complex indel profiles.

Conclusions:

CRISP-ID offers a powerful solution to the challenge of screening gene-edited clones with multiple indels.
The web application streamlines the analysis of gene editing outcomes, facilitating functional gene analysis.
This tool enhances the efficiency and reliability of genome engineering workflows.