[Zbrafish] Change from ZFNs to TALENs for future NIH-sponsored zebrafish gene knockout solicitations

Keith Joung via zbrafish%40net.bio.net (by keith.joung from zincfingers.org)
Sun Mar 11 18:09:32 EST 2012


Dear colleagues,

As many of you know, we have previously made two solicitations to the
community regarding the construction of engineered zinc finger pairs
for endogenous zebrafish gene targets.  As a result of these efforts,
we have constructed zinc finger pairs for sites in 49 different
zebrafish genes and have deposited these plasmids with the non-profit
plasmid distribution service Addgene (http://www.addgene.org/zfc/
arrays/).

We and others have recently shown that transcription activator-like
effector nucleases (TALENs) provide a robust alternative for
performing targeted gene knockout in zebrafish (Sander et al., Nat
Biotechnology 2011; Huang et al., Nat Biotechnology 2011).  The Joung
lab recently shown that the simplicity of TALEN design allows these
nucleases to be constructed in high-throughput using an automated
system we have developed and that the targeting range of TALENs is
essentially limitless (Reyon & Tsai et al., Nat Biotechnology, in
press).  Additional unpublished experiments from our groups have also
demonstrated that most TALENs induce mutations with remarkably high
efficiency and that this platform has a high success rate.

As a result of these recent advances in the field, , we plan to make
TALENs rather than ZFNs for future NIH-sponsored zebrafish gene
knockout solicitations to the community.  Doing so will enable more
precise alterations to be created within the gene due to the more
robust targeting range of TALENs and also improve the success rate.
Furthermore, because we have the ability to construct TALEN plasmids
in high-throughput, we will also be able to accelerate the number of
genes we target in each solicitation.

We hope that the availability of TALENs will be viewed as an exciting
advance for the community and welcome any comments you may have.

With best regards,

Keith Joung
Randy Peterson
Joanna Yeh
Jeffry Sander



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