Association for Biology Laboratory Education

Shhhhh...Keep It Down! Designing, Making, and Testing shRNA
 



Tested Studies in Laboratory Teaching, 2017, Volume 38

Alp Oran

Abstract

The following report is a modified description of an upper-level undergraduate lab course involving RNA interference (RNAi). RNAi is an umbrella term that includes any methodology that works to interfere with the expression of a target RNA sequence and is currently a major tool used by scientists around the world to understand gene function. A cost-effective PCR-based method for producing a popular form of RNAi, namely short hairpin RNA (shRNA), was adapted from the literature for the purposes of creating a mini project that undergraduate students with basic bench research skills would complete over a 12-week semester course. For the purposes of this communication and recognizing the varied needs of the readership, the author outlines different scenarios in which shRNA against any gene transcript can be created and tested as either a short independent lab module or as a core project for an entire lab course.

Keywords:  RNA interference, shRNA, RNAi, transfection, knockdown

University of Houston (2016)