Work

Using CRISPRi in Escherichia coli to emphasize experimental controls in a molecular microbiology laboratory

Year: 2025

Type: article

Abstract: ABSTRACT A CRISPR interference (CRISPRi) exercise was developed for an upper-level molecular microbiology laboratory to reinforce student skills in experimental design and controls. This CRISPRi knockdown method is a variation of the commonly used Streptococcus pyogenes Cas9 system and therefore relies on similar design and techniques. Students choose and design a CRISPRi target in Escherichia coli , clone the necessary tools, and test their system with spot plating and microscopy. The motivation for introducing this unit in a laboratory course was to help close gaps in students’ broader understanding of DNA and RNA structure, primer design, bacterial gene expression, and regulation. Once introduced, this exercise became a way to help students identify, design, and rationalize proper experimental controls.

Source: Journal of Microbiology and Biology Education

DOI: 10.1128/jmbe.00135-25

Author: Tammy J. Bullwinkle

Institution: The Ohio State University

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Quality education

Open Access status: gold

Estimated APC paid: $1,700