One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster
Summary: This one-step method generates, for any locus, a conditional knockout allele in Drosophila. The allele carries a bright fluorescent marker for easy screening and an attP landing site for subsequent genetic manipulations. After removing the selectable marker with Cre, the attP site can be us...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-06-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721002677 |
| id |
doaj-8652a259723149758a35c0d9c07f0b3d |
|---|---|
| record_format |
Article |
| spelling |
doaj-8652a259723149758a35c0d9c07f0b3d2021-06-21T04:25:42ZengElsevierSTAR Protocols2666-16672021-06-0122100560One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogasterJoyce J.S. Yu0Jean-Paul Vincent1Ian J. McGough2The Francis Crick Institute, London NW1 1AT, UKThe Francis Crick Institute, London NW1 1AT, UK; Corresponding authorThe Francis Crick Institute, London NW1 1AT, UK; Corresponding authorSummary: This one-step method generates, for any locus, a conditional knockout allele in Drosophila. The allele carries a bright fluorescent marker for easy screening and an attP landing site for subsequent genetic manipulations. After removing the selectable marker with Cre, the attP site can be used to insert DNA fragments expressing tagged or mutant alleles to determine protein localization and function in a tissue- and stage-specific manner. Only a single round of CRISPR-Cas9-mediated editing is required.For complete details on the use and execution of this protocol, please refer to the DWnt4[cKO] example in Yu et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166721002677CRISPRGeneticsModel Organisms |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Joyce J.S. Yu Jean-Paul Vincent Ian J. McGough |
| spellingShingle |
Joyce J.S. Yu Jean-Paul Vincent Ian J. McGough One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster STAR Protocols CRISPR Genetics Model Organisms |
| author_facet |
Joyce J.S. Yu Jean-Paul Vincent Ian J. McGough |
| author_sort |
Joyce J.S. Yu |
| title |
One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster |
| title_short |
One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster |
| title_full |
One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster |
| title_fullStr |
One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster |
| title_full_unstemmed |
One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster |
| title_sort |
one-step crispr-cas9 protocol for the generation of plug & play conditional knockouts in drosophila melanogaster |
| publisher |
Elsevier |
| series |
STAR Protocols |
| issn |
2666-1667 |
| publishDate |
2021-06-01 |
| description |
Summary: This one-step method generates, for any locus, a conditional knockout allele in Drosophila. The allele carries a bright fluorescent marker for easy screening and an attP landing site for subsequent genetic manipulations. After removing the selectable marker with Cre, the attP site can be used to insert DNA fragments expressing tagged or mutant alleles to determine protein localization and function in a tissue- and stage-specific manner. Only a single round of CRISPR-Cas9-mediated editing is required.For complete details on the use and execution of this protocol, please refer to the DWnt4[cKO] example in Yu et al. (2020). |
| topic |
CRISPR Genetics Model Organisms |
| url |
http://www.sciencedirect.com/science/article/pii/S2666166721002677 |
| work_keys_str_mv |
AT joycejsyu onestepcrisprcas9protocolforthegenerationofplugampplayconditionalknockoutsindrosophilamelanogaster AT jeanpaulvincent onestepcrisprcas9protocolforthegenerationofplugampplayconditionalknockoutsindrosophilamelanogaster AT ianjmcgough onestepcrisprcas9protocolforthegenerationofplugampplayconditionalknockoutsindrosophilamelanogaster |
| _version_ |
1721368889518981120 |