| RRC ID |
48897
|
| 著者 |
Adame V, Chapapas H, Cisneros M, Deaton C, Deichmann S, Gadek C, Lovato TL, Chechenova MB, Guerin P, Cripps RM.
|
| タイトル |
An undergraduate laboratory class using CRISPR/Cas9 technology to mutate drosophila genes.
|
| ジャーナル |
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.
|
| 巻・号 |
44(3)
|
| ページ |
263-75
|
| 公開日 |
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
|
| 引用数 |
15
|
|
WOS 分野
|
EDUCATION, SCIENTIFIC DISCIPLINES
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ショウジョウバエ |
CAS-0002 |
