RRC ID 48897
Author Adame V, Chapapas H, Cisneros M, Deaton C, Deichmann S, Gadek C, Lovato TL, Chechenova MB, Guerin P, Cripps RM.
Title An undergraduate laboratory class using CRISPR/Cas9 technology to mutate drosophila genes.
Journal Biochem Mol Biol Educ
Abstract CRISPR/Cas9 genome editing technology is used in the manipulation of genome sequences and gene expression. Because of the ease and rapidity with which genes can be mutated using CRISPR/Cas9, we sought to determine if a single-semester undergraduate class could be successfully taught, wherein students isolate mutants for specific genes using CRISPR/Cas9. Six students were each assigned a single Drosophila gene, for which no mutants currently exist. Each student designed and created plasmids to encode single guide RNAs that target their selected gene; injected the plasmids into Cas9-expressing embryos, in order to delete the selected gene; carried out a three-generation cross to test for germline transmission of a mutated allele and generate a stable stock of the mutant; and characterized the mutant alleles by PCR and sequencing. Three genes out of six were successfully mutated. Pre- and post- survey evaluations of the students in the class revealed that student attitudes towards their research competencies increased, although the changes were not statistically significant. We conclude that it is feasible to develop a laboratory genome editing class, to provide effective laboratory training to undergraduate students, and to generate mutant lines for use by the broader scientific community. © 2016 by The International Union of Biochemistry and Molecular Biology, 44:263-275, 2016.
Volume 44(3)
Pages 263-75
Published 2016-5-6
DOI 10.1002/bmb.20950
PMID 27009801
PMC PMC5377917
MeSH Amino Acid Sequence Animals Base Sequence CRISPR-Cas Systems / genetics* Drosophila / genetics* Drosophila Proteins / genetics* Female Gene Targeting / methods* Genetic Engineering Genome, Insect Male Molecular Biology / education* Mutation / genetics Plasmids / genetics RNA Editing / genetics*
IF 0.924
Times Cited 15
Drosophila CAS-0002