[Laboratory design and practice for undergraduates: Using RNAi to modulate gene expression].

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• RNA interference (RNAi) is a crucial gene silencing mechanism utilizing double-stranded RNAs.
• RNAi has become a vital tool for reverse genetics research.
• Effective pedagogical approaches are needed to teach RNAi principles and techniques.

Purpose of the Study:

• To enhance undergraduate understanding of RNA interference principles and technology.
• To develop practical laboratory skills in gene silencing experiments.
• To provide hands-on experience in designing and validating small interfering RNAs (siRNAs).

Main Methods:

• Students selected target genes, designed siRNAs and primers, and performed gene silencing experiments.
• Experiments involved transfecting 3T3-L1 cells with siRNAs targeting Mus musculus acyl-CoA synthetase long-chain family member 1 (Acsl1) via electroporation.
• Messenger RNA (mRNA) expression levels were quantified using relative quantitative PCR after cDNA synthesis.

Main Results:

• Both designed siRNA pairs demonstrated over 60% gene silencing efficiency for Acsl1.
• Approximately 83% of students successfully completed the experiments over three years.
• Students consistently identified at least one effective siRNA pair.

Conclusions:

• The implemented teaching strategy effectively improves students' grasp of RNAi.
• This practical approach enhances students' laboratory skills and scientific inquiry abilities.
• The methodology provides a robust framework for teaching gene silencing techniques in undergraduate education.