[Zbrafish] Re: FINAL Update on Second Solicitation of the NIH-sponsored zebrafish ZFN project

Keith Joung via zbrafish%40net.bio.net (by keith.joung from zincfingers.org)
Sat Apr 28 07:44:08 EST 2012


A quick follow-up message to let you know that all of the TALENs and
ZFNs made for the second solicitation of the NIH-sponsored zebrafish
targeted nuclease project are now available from Addgene:

For ZFNs:  http://www.addgene.org/zfc/arrays/

For TALENs:  http://www.addgene.org/talengineering/TALENzebrafish/

A third solicitation round for TALENs targeted to zebrafish genes will
be announced in the coming weeks.

Best,

Keith

On Mar 11, 7:10 pm, Keith Joung <keith.jo... from zincfingers.org> wrote:
> Dear all,
>
> We write to provide a final update on the status of our second effort
> to target 48 zebrafish genes requested by the community.  We have now
> successfully generated targeting constructs for 29 of the 48 genes, as
> outlined below, and are preparing to release another solicitation for
> additional genes to be targeted.
>
> In our last update from October of last year, we initially identified
> zinc finger array pairs made by OPEN or CoDA that can be used to
> modify 13 different endogenous zebrafish genes.  We also reported that
> we knew with certainty that we were unable to obtain zinc finger array
> pairs for 10 zebrafish genes.
>
> We have several updates to report:
>
> (1) We discovered a clerical error after our last update e-mail and
> realized that we had misidentified 8 of the 13 gene targets we
> originally reported success for.  The five correctly-identified genes
> remain in the Group I list below and are preceded by an asterisk (*).
>
> (2) For the eight genes for which we originally reported success but
> actually did NOT obtain zinc finger array pairs, we have constructed
> TALE nucleases (TALENs) targeted to sites chosen in consultation with
> the original requestors.  As shown by our group and others, TALENs can
> also be used to introduce targeted alterations in zebrafish.  The
> names of these eight genes are shown in the Group III list below.
>
> (3) We identified additional CoDA and/or OPEN ZF pairs for sites in
> sixteen more genes (including four genes previously reported in Group
> II, now preceded by a pound sign (#) in the Group I list below).
>
> (4) We confirmed that we were unable to obtain CoDA ZF pairs for sites
> in 9 additional genes (these are listed in Group II and preceded by a
> double asterisk (**)).
>
> We thank you for your patience with this process.  Note that all of
> the plasmids encoding the 21 pairs of ZFs from Group I are available
> now from Addgene:  http://www.addgene.org/zfc/arrays/.  The eight
> pairs of TALENs from Group III will also be available from Addgene in
> the coming weeks here:  http://www.addgene.org/talengineering/TALENzebrafish/.
>
> We plan to make another solicitation for target genes in the coming
> month.
>
> With best regards,
>
> Keith Joung
> Randy Peterson
> Joanna Yeh
> Jeffry Sander
> Maureen Regan
>
> Group I genes (ZF array pairs successfully obtained): (21 genes)
>
> *gene_name "LOC566040"transcript_name "LOC566040"
> *gene_name "esrrga"transcript_name "esrrga"
> *gene_name "HACE1"
> *gene_name "atp2c1"transcript_name "atp2c1"
> *gene_name "dio2"transcript_name "dio2"
> gene_name "im:7163548"transcript_name "im:7163548"
> gene_name "fam82a2"transcript_name "fam82a2"
> gene_name "morc3b"transcript_name "NP_001003579.1"
> gene_name "psen2"transcript_name "psen2"
> gene_name "zgc:162148"transcript_name "MICU1_DANRE"
> gene_name "unc119b"transcript_name "unc119b"
> #gene_name "cx52.9"transcript_name "cx52.9"
> gene_name "flt1"transcript_name "A4QNT7_DANRE"
> #gene_name "yap1"transcript_name "yap1"
> gene_name "pde3b"transcript_name "pde3b"
> gene_name "col6a2"transcript_name "col6a2"
> #gene_name "SDC3"
> gene_name "zgc:111821"transcript_name "zgc:111821"
> gene_name "zgc:65890"transcript_name "zgc:65890"
> gene_name "panx2"transcript_name "B1P2E8_DANRE"
> #gene_name "si:ch211-221n23.1"transcript_name "ZN238_DANRE"
>
> Group II genes (failed to obtain ZF array pairs) (19 genes):
>
> gene_name "hnf4a"transcript_name "NP_919349.1"
> gene_name "wt1b"transcript_name "wt1b"
> gene_name "jup"transcript_name "jup"
> gene_name "myca"transcript_name "myca"
> gene_name "igf2bp1"transcript_name "igf2bp1"
> gene_name "jazf1a"
> **gene_name "emp2"transcript_name "emp2"
> **gene_name "mych"transcript_name "mych"
> **gene_name "sqstm1"transcript_name "NP_998338.1"
> **gene_name "ahrrb"transcript_name "ahrrb"
> **gene_name A9JTD0_DANREtranscript_name A9JTD0_DANRE
> **gene_name "f9"transcript_name "NM_182868.1"
> **gene_name "coq6"transcript_name "NP_001038869.1"
> **gene_name "csf3r"transcript_name "csf3r"
> **gene_name "trpv1"transcript_name "B6RTA2_DANRE"
> **gene_name "cadm2a"transcript_name "cadm2a"
> **gene_name "dab1a"transcript_name "A2BEF7_DANRE"
> **gene_name "LOC565975"transcript_name "LOC565975"
> **gene_name "vasa"transcript_name "vasa"
>
> Group III genes (for which we have made TALENs) (8 genes):
>
> gene_name "ptp4a3"transcript_name "ptp4a3"
> gene_name "mchr1b"transcript_name "mchr1b"
> gene_name "arsa"transcript_name "arsa"
> gene_name "park2"transcript_name "park2"
> gene_name "v2rl1"transcript_name "Q4VDB3_DANRE"
> gene_name "gabra5"transcript_name "gabra5"
> gene_name "drd4a"transcript_name "drd4a"
> gene_name "gdf2"transcript_name "gdf2"




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