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In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

In vitro Mutagenesis

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Related Experiment Video

Updated: Jul 25, 2026

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Published on: December 23, 2022

Development of Cas13a-based Assays for

Lao-Tzu Allan-Blitz^1,2,3, Palak Shah^2,3, Gordon Adams^2,3

¹Division of Global Health Equity: Department of Medicine, Brigham and Women's Hospital, Boston, MA.

Medrxiv : the Preprint Server for Health Sciences

|June 9, 2023

Summary

New CRISPR-based SHERLOCK assays can detect Neisseria gonorrhoeae and predict ciprofloxacin resistance. This offers a low-cost diagnostic tool for sexually transmitted infections, especially in resource-limited settings.

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Published on: April 25, 2025

Area of Science:

Molecular Diagnostics
Antimicrobial Resistance
CRISPR Technology

Background:

Neisseria gonorrhoeae is a common sexually transmitted infection with rising antimicrobial resistance.
Current diagnostic methods are costly and require extensive lab infrastructure, limiting use in low-resource areas.
CRISPR-based diagnostics like SHERLOCK offer potential for low-cost pathogen and resistance detection.

Approach:

Developed Specific High-sensitivity Enzymatic Reporter unLOCKing (SHERLOCK) assays using CRISPR-Cas13a.
Designed RNA guides and primers to detect N. gonorrhoeae via the porA gene.
Created assays to predict ciprofloxacin susceptibility by targeting a gyrA gene mutation.

Key Points:

SHERLOCK assays accurately detected N. gonorrhoeae in 14 isolates without cross-reactivity.
Fluorescence-based assay correctly distinguished ciprofloxacin-resistant from susceptible N. gonorrhoeae isolates (20 vs. 3).
Genotype predictions for ciprofloxacin resistance showed 100% concordance with DNA sequencing.

Conclusions:

Successfully developed Cas13a-based SHERLOCK assays for N. gonorrhoeae detection.
These assays can differentiate ciprofloxacin-resistant from susceptible strains.
This technology holds promise for accessible diagnostics of gonorrhea and its resistance patterns.