Please use this identifier to cite or link to this item: https://doi.org/10.1007/s10126-013-9556-6
DC FieldValue
dc.titleParameters and Efficiency of Direct Gene Disruption by Zinc Finger Nucleases in Medaka Embryos
dc.contributor.authorZhang, X.
dc.contributor.authorGuan, G.
dc.contributor.authorChen, J.
dc.contributor.authorNaruse, K.
dc.contributor.authorHong, Y.
dc.date.accessioned2014-10-27T08:36:13Z
dc.date.available2014-10-27T08:36:13Z
dc.date.issued2014-04
dc.identifier.citationZhang, X., Guan, G., Chen, J., Naruse, K., Hong, Y. (2014-04). Parameters and Efficiency of Direct Gene Disruption by Zinc Finger Nucleases in Medaka Embryos. Marine Biotechnology 16 (2) : 125-134. ScholarBank@NUS Repository. https://doi.org/10.1007/s10126-013-9556-6
dc.identifier.issn14362228
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/101348
dc.description.abstractZinc finger nucleases (ZFNs) can generate targeted gene disruption (GD) directly in developing embryos of zebrafish, mouse and human. In the fish medaka, ZFNs have been attempted on a transgene. Here, we developed procedures and parameters for ZFN-mediated direct GD on the gonad-specifically expressed gsdf locus in medaka. A pair of ZFNs was designed to target the first exon of gsdf and their synthetic mRNAs were microinjected into 1-cell stage embryos. We reveal dose-dependent survival rate and GD efficiency. In fry, ZFN mRNA injection at 10 ng/μl led to a GD efficiency of 30 %. This value increased up to nearly 100 % when the dose was enhanced to 40 ng/μl. In a typical series of experiments of ZFN mRNA injection at 10 ng/μl, 420 injected embryos developed into 94 adults, 4 of which had altered gsdf alleles. This leads to a GD efficacy of ~4 % in the adulthood. Sequencing revealed a wide variety of subtle allelic alterations including additions and deletions of 1~18 bp in length in ZFN-injected samples. Most importantly, one of the 4 adults examined was capable of germline transmission to 15.2 % of its F1 progeny. Interestingly, ontogenic analyses of the allelic profile revealed that GD commenced early in development, continued during subsequent stages of development and in primordia for different adult organs of the three germ layers. These results demonstrate the feasibility and-for the first time to our knowledge-the efficacy of ZFN-mediated direct GD on a chromosomal gene in medaka embryos. © 2013 Springer Science+Business Media New York.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1007/s10126-013-9556-6
dc.sourceScopus
dc.subjectGene disruption
dc.subjectGene targeting
dc.subjectGenome editing
dc.subjectgsdf
dc.subjectZFN
dc.subjectZinc finger nuclease
dc.typeArticle
dc.contributor.departmentBIOLOGICAL SCIENCES
dc.description.doi10.1007/s10126-013-9556-6
dc.description.sourcetitleMarine Biotechnology
dc.description.volume16
dc.description.issue2
dc.description.page125-134
dc.description.codenMABIF
dc.identifier.isiut000331652200001
Appears in Collections:Staff Publications

Show simple item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

13
checked on Nov 30, 2021

WEB OF SCIENCETM
Citations

8
checked on Nov 30, 2021

Page view(s)

187
checked on Dec 2, 2021

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.