CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish
Here we present a protocol for the conversion of eGFP-transgenic zebrafish lines into lines expressing Gal4 from the same locus. This conversion allows the in-depth analysis of the former eGFP-expressing cell population; with the Gal4-upstream activating sequence (UAS) system, diverse UAS transgenes...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
13 November 2014
|
| In: |
Nature protocols
Year: 2014, Volume: 9, Issue: 12, Pages: 2823-2840 |
| ISSN: | 1750-2799 |
| DOI: | 10.1038/nprot.2014.187 |
| Online Access: | Resolving-System, lizenzpflichtig, Volltext: https://doi.org/10.1038/nprot.2014.187 Verlag, lizenzpflichtig, Volltext: https://www.nature.com/articles/nprot.2014.187 
|
| Author Notes: | Thomas O. Auer, Karine Duroure, Jean-Paul Concordet, Filippo Del Bene |
MARC
| LEADER | 00000caa a22000002c 4500 | ||
|---|---|---|---|
| 001 | 1735546828 | ||
| 003 | DE-627 | ||
| 005 | 20220818224807.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 201014s2014 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1038/nprot.2014.187 |2 doi | |
| 035 | |a (DE-627)1735546828 | ||
| 035 | |a (DE-599)KXP1735546828 | ||
| 035 | |a (OCoLC)1341371290 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 32 |2 sdnb | ||
| 100 | 1 | |a Auer, Thomas |e VerfasserIn |0 (DE-588)1065633084 |0 (DE-627)816265879 |0 (DE-576)425314065 |4 aut | |
| 245 | 1 | 0 | |a CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish |c Thomas O. Auer, Karine Duroure, Jean-Paul Concordet, Filippo Del Bene |
| 246 | 3 | 0 | |a eGFP Gal4 - |
| 264 | 1 | |c 13 November 2014 | |
| 300 | |a 18 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Gesehen am 14.10.2020 | ||
| 520 | |a Here we present a protocol for the conversion of eGFP-transgenic zebrafish lines into lines expressing Gal4 from the same locus. This conversion allows the in-depth analysis of the former eGFP-expressing cell population; with the Gal4-upstream activating sequence (UAS) system, diverse UAS transgenes can be transactivated. Site-specific targeting of the gene encoding eGFP is achieved using the clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) system. A single-guide RNA (sgRNA) that targets eGFP is injected into embryos together with a donor vector containing an optimized version of Gal4 (KalTA4) to trigger integration of the donor into the targeted eGFP genomic location. To enable screening for successful integration events, injection is performed in a UAS:RFP transgenic background; fish showing mosaic eGFP-to-RFP conversion are raised to adulthood. The progeny of these adult fish are then screened for stable germline transmission, and converted progeny are used to generate stable lines. We have been able to generate two stably converted transgenic lines within 4 months. | ||
| 700 | 1 | |a Duroure, Karine |e VerfasserIn |4 aut | |
| 700 | 1 | |a Concordet, Jean-Paul |e VerfasserIn |4 aut | |
| 700 | 1 | |a Del Bene, Filippo |e VerfasserIn |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Nature protocols |d Basingstoke : Nature Publishing Group, 2006 |g 9(2014), 12, Seite 2823-2840 |h Online-Ressource |w (DE-627)515980552 |w (DE-600)2244966-8 |w (DE-576)276000455 |x 1750-2799 |7 nnas |a CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish |
| 773 | 1 | 8 | |g volume:9 |g year:2014 |g number:12 |g pages:2823-2840 |g extent:18 |a CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish |
| 856 | 4 | 0 | |u https://doi.org/10.1038/nprot.2014.187 |x Resolving-System |x Verlag |z lizenzpflichtig |3 Volltext |
| 856 | 4 | 0 | |u https://www.nature.com/articles/nprot.2014.187 |x Verlag |z lizenzpflichtig |3 Volltext |
| 951 | |a AR | ||
| 992 | |a 20201014 | ||
| 993 | |a Article | ||
| 994 | |a 2014 | ||
| 998 | |g 1065633084 |a Auer, Thomas |m 1065633084:Auer, Thomas |d 140000 |e 140000PA1065633084 |k 0/140000/ |p 1 |x j | ||
| 999 | |a KXP-PPN1735546828 |e 3780002671 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"person":[{"family":"Auer","given":"Thomas","display":"Auer, Thomas","roleDisplay":"VerfasserIn","role":"aut"},{"family":"Duroure","given":"Karine","roleDisplay":"VerfasserIn","display":"Duroure, Karine","role":"aut"},{"display":"Concordet, Jean-Paul","roleDisplay":"VerfasserIn","role":"aut","family":"Concordet","given":"Jean-Paul"},{"display":"Del Bene, Filippo","roleDisplay":"VerfasserIn","role":"aut","family":"Del Bene","given":"Filippo"}],"title":[{"title":"CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish","title_sort":"CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafish"}],"type":{"bibl":"article-journal","media":"Online-Ressource"},"note":["Gesehen am 14.10.2020"],"language":["eng"],"recId":"1735546828","name":{"displayForm":["Thomas O. Auer, Karine Duroure, Jean-Paul Concordet, Filippo Del Bene"]},"origin":[{"dateIssuedKey":"2014","dateIssuedDisp":"13 November 2014"}],"id":{"eki":["1735546828"],"doi":["10.1038/nprot.2014.187"]},"physDesc":[{"extent":"18 S."}],"relHost":[{"origin":[{"dateIssuedDisp":"2006-","publisher":"Nature Publishing Group","dateIssuedKey":"2006","publisherPlace":"Basingstoke"}],"id":{"issn":["1750-2799"],"zdb":["2244966-8"],"eki":["515980552"]},"physDesc":[{"extent":"Online-Ressource"}],"title":[{"title":"Nature protocols","title_sort":"Nature protocols"}],"pubHistory":["1.2006 -"],"titleAlt":[{"title":"Protocols"}],"part":{"pages":"2823-2840","issue":"12","year":"2014","extent":"18","volume":"9","text":"9(2014), 12, Seite 2823-2840"},"type":{"bibl":"periodical","media":"Online-Ressource"},"disp":"CRISPR/Cas9-mediated conversion of eGFP- into Gal4-transgenic lines in zebrafishNature protocols","note":["Gesehen am 18. September 2018"],"language":["eng"],"recId":"515980552"}]} | ||
| SRT | |a AUERTHOMASCRISPRCAS91320 | ||