Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases

Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases Direct genomic manipulation at a specific locus is still not feasible in most vertebrate model organisms. In vertebrate cell lines, genomic lesions at a specific site have been introduced using zinc-finger nucleases (ZFNs) . Here we adapt this technology to create targeted mutations in the zebrafish germ line. ZFNs were engineered that recognize sequences in the zebrafish ortholog of the vascular endothelial growth factor-2 receptor, kdr (also known as kdra ). Co-injection of mRNAs encoding these ZFNs into one-cell-stage zebrafish embryos led to mutagenic lesions at the target site that were transmitted through the germ line with high frequency. The use of engineered ZFNs to introduce heritable mutations into a genome obviates the need for embryonic stem cell lines and should be applicable to most animal species for which early-stage embryos are easily accessible. The ability to perform targeted genomic manipulation in the mouse using embryonic stem cell lines has established it as a central vertebrate model system . Similar manipulations in other vertebrate models have largely failed owing to the difficulty of generating embryonic stem cell lines. ZFNs, which are a chimeric fusion between a Cys 2 His 2 zinc-finger protein (ZFP) and the cleavage domain of http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Biotechnology Springer Journals

Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases

Loading next page...
 
/lp/springer-journals/targeted-gene-inactivation-in-zebrafish-using-engineered-zinc-finger-FS5wLvpPGa
Publisher
Springer Journals
Copyright
Copyright © 2008 Nature Publishing Group
ISSN
1087-0156
eISSN
1087-0156
D.O.I.
10.1038/nbt1398
Publisher site
See Article on Publisher Site

Abstract

Direct genomic manipulation at a specific locus is still not feasible in most vertebrate model organisms. In vertebrate cell lines, genomic lesions at a specific site have been introduced using zinc-finger nucleases (ZFNs) . Here we adapt this technology to create targeted mutations in the zebrafish germ line. ZFNs were engineered that recognize sequences in the zebrafish ortholog of the vascular endothelial growth factor-2 receptor, kdr (also known as kdra ). Co-injection of mRNAs encoding these ZFNs into one-cell-stage zebrafish embryos led to mutagenic lesions at the target site that were transmitted through the germ line with high frequency. The use of engineered ZFNs to introduce heritable mutations into a genome obviates the need for embryonic stem cell lines and should be applicable to most animal species for which early-stage embryos are easily accessible. The ability to perform targeted genomic manipulation in the mouse using embryonic stem cell lines has established it as a central vertebrate model system . Similar manipulations in other vertebrate models have largely failed owing to the difficulty of generating embryonic stem cell lines. ZFNs, which are a chimeric fusion between a Cys 2 His 2 zinc-finger protein (ZFP) and the cleavage domain of

Journal

Nature BiotechnologySpringer Journals

Published: May 25, 2008

There are no references for this article.

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off