title: An undergraduate laboratory class using CRISPR/Cas9 technology to mutate drosophila genes.
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.
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Adame
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Holly
authors.initials: H
authors.lastname: Chapapas
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Marilyn
authors.initials: M
authors.lastname: Cisneros
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Carol
authors.initials: C
authors.lastname: Deaton
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Sophia
authors.initials: S
authors.lastname: Deichmann
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Chauncey
authors.initials: C
authors.lastname: Gadek
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: TyAnna L
authors.initials: TL
authors.lastname: Lovato
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Maria B
authors.initials: MB
authors.lastname: Chechenova
authors.affiliation: Institute for Social Research, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Paul
authors.initials: P
authors.lastname: Guerin
authors.affiliation: Department of Biology, University of New Mexico, Albuquerque, New Mexico, 87131.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Cripps
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D005260:Female; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering; D049750:Genome, Insect; D008297:Male; D008967:Molecular Biology / Q000193:education*; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D017393:RNA Editing / Q000235:genetics*
chemical_list: D029721:Drosophila Proteins
keywords: CRISPR; drosophila; laboratory class; research; undergraduate