Fluorescently labeled TracrRNA improves work flow and facilitates successful genome editing in zebrafish

Research output: Contribution to journalArticleOtherpeer-review

Abstract

Gene editing using clustered regularly interspaced short palindromic repeats (CRISPR) is widely used throughout the zebrafish community for the generation of knockouts and knockins. One of the bottlenecks that exists during the process is the laborious screening of injected embryos for F0 founder fish or CRISPants, weeks after the injection date. In this study we show that the use of fluorescently tagged tracrRNA and sorting for fluorescent embryos as early as the 512-cell stage using stereomicroscope significantly improve yield of fish with successfully CRISPR/Cas9-edited genomes. This is a cost-effective strategy that significantly improves workflow and efficacy in genome editing in particular for less experienced researchers.

Original languageEnglish
Pages (from-to)135-137
Number of pages3
JournalZebrafish
Volume16
Issue number1
DOIs
Publication statusPublished - 31 Jan 2019

Keywords

  • CRISPR
  • knockin
  • knockout
  • loss-of-function
  • mutant
  • teleost

Cite this

@article{edd8a9ec70ae44699b0c7ee57625aab1,
title = "Fluorescently labeled TracrRNA improves work flow and facilitates successful genome editing in zebrafish",
abstract = "Gene editing using clustered regularly interspaced short palindromic repeats (CRISPR) is widely used throughout the zebrafish community for the generation of knockouts and knockins. One of the bottlenecks that exists during the process is the laborious screening of injected embryos for F0 founder fish or CRISPants, weeks after the injection date. In this study we show that the use of fluorescently tagged tracrRNA and sorting for fluorescent embryos as early as the 512-cell stage using stereomicroscope significantly improve yield of fish with successfully CRISPR/Cas9-edited genomes. This is a cost-effective strategy that significantly improves workflow and efficacy in genome editing in particular for less experienced researchers.",
keywords = "CRISPR, knockin, knockout, loss-of-function, mutant, teleost",
author = "Mustafa Hamimi and Mitra Khabooshan and Castillo, {Hozana A.} and Jan Kaslin",
year = "2019",
month = "1",
day = "31",
doi = "10.1089/zeb.2018.1669",
language = "English",
volume = "16",
pages = "135--137",
journal = "Zebrafish",
issn = "1545-8547",
publisher = "Mary Ann Liebert Inc",
number = "1",

}

Fluorescently labeled TracrRNA improves work flow and facilitates successful genome editing in zebrafish. / Hamimi, Mustafa; Khabooshan, Mitra; Castillo, Hozana A.; Kaslin, Jan.

In: Zebrafish, Vol. 16, No. 1, 31.01.2019, p. 135-137.

Research output: Contribution to journalArticleOtherpeer-review

TY - JOUR

T1 - Fluorescently labeled TracrRNA improves work flow and facilitates successful genome editing in zebrafish

AU - Hamimi, Mustafa

AU - Khabooshan, Mitra

AU - Castillo, Hozana A.

AU - Kaslin, Jan

PY - 2019/1/31

Y1 - 2019/1/31

N2 - Gene editing using clustered regularly interspaced short palindromic repeats (CRISPR) is widely used throughout the zebrafish community for the generation of knockouts and knockins. One of the bottlenecks that exists during the process is the laborious screening of injected embryos for F0 founder fish or CRISPants, weeks after the injection date. In this study we show that the use of fluorescently tagged tracrRNA and sorting for fluorescent embryos as early as the 512-cell stage using stereomicroscope significantly improve yield of fish with successfully CRISPR/Cas9-edited genomes. This is a cost-effective strategy that significantly improves workflow and efficacy in genome editing in particular for less experienced researchers.

AB - Gene editing using clustered regularly interspaced short palindromic repeats (CRISPR) is widely used throughout the zebrafish community for the generation of knockouts and knockins. One of the bottlenecks that exists during the process is the laborious screening of injected embryos for F0 founder fish or CRISPants, weeks after the injection date. In this study we show that the use of fluorescently tagged tracrRNA and sorting for fluorescent embryos as early as the 512-cell stage using stereomicroscope significantly improve yield of fish with successfully CRISPR/Cas9-edited genomes. This is a cost-effective strategy that significantly improves workflow and efficacy in genome editing in particular for less experienced researchers.

KW - CRISPR

KW - knockin

KW - knockout

KW - loss-of-function

KW - mutant

KW - teleost

UR - http://www.scopus.com/inward/record.url?scp=85060920639&partnerID=8YFLogxK

U2 - 10.1089/zeb.2018.1669

DO - 10.1089/zeb.2018.1669

M3 - Article

VL - 16

SP - 135

EP - 137

JO - Zebrafish

JF - Zebrafish

SN - 1545-8547

IS - 1

ER -