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 language | English |
|---|---|
| Pages (from-to) | 135-137 |
| Number of pages | 3 |
| Journal | Zebrafish |
| Volume | 16 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 31 Jan 2019 |
Keywords
- CRISPR
- knockin
- knockout
- loss-of-function
- mutant
- teleost
Cite this
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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 journal › Article › Other › peer-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 -