Dear Colleagues,

As many of you know, I submitted a proposal to the NIH in January to support
high-throughput production of gene knockout strains in Tetrahymena
thermophila.  The reviewers strongly appreciated the importance of this
resource to the community, but preferred a different strategy – essentially,
they thought generating gene deletions would be preferable to insertional
mutations (which I originally favored as being more high throughput).  They
also recommended incorporating “barcode” sequences at each KO position that
would allow for competitive growth experiments on pools of KO strains,
similar to what has been done in yeast (e.g. Science 320: 362-365).  I have
come to agree with the reviewers and am preparing a resubmission of the
proposal for a deadline of September 25th.  The central aim of the proposal
will be the same as the first – to produce germline knockouts of genes
selected by their interest to members of the research community.  The exact
number of KOs I will aim for will depend on budget constraints that are not
yet worked out, but it will be between 1000 and 2000.  They will be made
available to all through the Tetrahymena Stock Center.

The many letters I received for the first submission were instrumental in
convincing reviewers of the high level of community support for this
resource.  If you wrote one of those letters, could you please send me an
updated version ASAP?!  If you are interested in using the powerful barcode
strategy to identify genes of interest, please include that in your letter.
 Also, please update me on recent progress in your lab, and any changes in
your research priorities that affect the kinds of genes you are interested
in obtaining knockouts for.  If you have any questions, or if you didn’t
send a letter previously but have an interest in this resource, please
contact me ASAP.  With the changes I am making and your continued support, I
am optimistic that the resubmission will be funded.

Thanks,
Bob Coyne

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