ERIC Number: EJ1462631
Record Type: Journal
Publication Date: 2025-Mar
Pages: 10
Abstractor: As Provided
ISBN: N/A
ISSN: ISSN-1470-8175
EISSN: EISSN-1539-3429
Available Date: 2024-12-19
Practical Laboratory Class to Assess Gene Silencing Using CRISPR Interference (CRISPRi) Technology in the Archaeon "Haloferax volcanii"
Biochemistry and Molecular Biology Education, v53 n2 p155-164 2025
Perturbation of gene expression using RNA interference (RNAi) or CRISPR interference (CRISPRi) is a useful strategy to explore the function of essential genes. In the archaeon "Haloferax volcanii," the CRISPR-Cas system has been adapted as a CRISPRi tool to silence the expression of specific genes. We developed a laboratory class (LC) to conceptualize gene silencing through inactivation of the "H. volcanii" LonB protease gene, a negative regulator of carotenoid pigments biosynthesis, using CRISPRi. This LC has been successfully applied in the Biology and Biochemistry of Microorganisms course for undergraduate students of Biology in 2022 and 2023. The following objectives were proposed: (a) generate "H. volcanii" mutant strains with reduced expression of the "lonB" gene using CRISPRi; (b) examine the effect of "lonB" gene silencing on cell pigmentation and growth rate; (c) assess "lonB" gene repression by Western blotting (WB). This LC allows students to obtain and screen CRISPRi silenced-mutants by means of simple procedures using a non-pathogenic organism as well as handle basic microbiology, biochemistry and molecular biology protocols. Additionally, the LC fosters social actions through collaborative work (experimental work), the interpretation and discussion of data and the ability to communicate outcomes orally and in a written format (scientific report).
Descriptors: Undergraduate Students, Laboratory Experiments, Science Instruction, Biochemistry, Molecular Biology, Molecular Structure, Microbiology, Computer Uses in Education, Cytology, Cooperative Learning, Communication Skills, Research Reports
Wiley. Available from: John Wiley & Sons, Inc. 111 River Street, Hoboken, NJ 07030. Tel: 800-835-6770; e-mail: cs-journals@wiley.com; Web site: https://www-wiley-com.bibliotheek.ehb.be/en-us
Publication Type: Journal Articles; Reports - Evaluative
Education Level: Higher Education; Postsecondary Education
Audience: N/A
Language: English
Sponsor: N/A
Authoring Institution: N/A
Grant or Contract Numbers: N/A
Author Affiliations: 1Instituto de Investigaciones Biológicas (IIB-CONICET-UNMDP), Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata, Mar del Plata, Argentina; 2Biology II, Ulm University, Ulm, Germany