Chlamy Mutant Library Project

 

Overview

We are generating a genome-wide collection of mutants with known mutation sites in Chlamydomonas.

This is the first such collection in any single-celled photosynthetic eukaryote.

The goal of this NSF-funded project is to isolate two mutants for 80% of all genes. We are leveraging novel high-throughput insertion site mapping and robotic mutant propagation strategies developed in our lab. The mutants will be distributed to the community through the Chlamydomonas Resource Center.

The team

The PI on the project is Martin Jonikas. Other PI's involved in the project are Arthur Grossman at Carnegie (Co-PI on the grant, advising on all aspects of the project), Pete Lefebvre at the University of Minnesota (who will oversee distribution of the resource to the community), and Eva Huala at Carnegie (developing the mutant distribution website).

The daily operations of the project are led by Xiaobo Li. Also working on the project are: Weronika Patena, Ru Zhang, Spencer Gang, Sean Blum, Nina Ivanova, Rebecca Yue.

Early distribution - only through 9pm US Pacific time on June 23rd, 2014 (hard deadline)

We presently have a pilot collection, containing ~1,700 strains (covering nearly 10% of all genes) where we have a high confidence in the insertion site. We aim to publish this work, but a substantial amount of validation and analysis remains to be done. We have received a large number of requests for collaborative sharing of specific individual mutants before we publish. We want to help the community, but at the same time, we need to keep our limited resources focused on our science (we are a young lab). Additionally, rather than sharing the mutants with a select few collaborators, we thought it would be fairest to make it possible for any lab in the community to collaborate with us and receive a mutant. To meet these goals, we have come up with a scheme to efficiently distribute the most desired mutants with relatively little impact on our science (we realize it's a bit impersonal, but it's the best we could do).

Download the list of available mutants by clicking this text.

Important notes:

a. Due to our limited manpower, we can only send out at most 1 mutant per laboratory Principal Investigator. If you received a mutant during the previous distribution, we can't send you another unless you email us with evidence that the insertion site in the original mutant was incorrect.
b. In order to receive your mutant, you must submit your request form by 9pm on June 23rd, 2014. We will strive to send out the mutants as soon as possible (we are aiming for the end of June).
c. For logistical reasons, we can only ship to places where FedEx ships; and are asking the requester to pay for shipment with their FedEx account.
d. The mutant we send may not be what we think it is. There is a 25% chance that the mutant we send does not have an insertion in the gene that we indicated (due to the messiness of insertion sites in Chlamy). You are responsible for verifying the flanking sequence on both sides of the annotated insertion site by PCR and sequencing, and for verifying the disruption of the wild-type locus. If your mutant is not what you expected, we unfortunately won't have the bandwidth to help you, nor will we be able to send you a different mutant.
e. If the strain dies, we won't be able to send you another one. So please propagate it carefully.
f. The identities of the mutants sent out and the labs that received them will be public information. Here is the list of strains distributed to date.

What we will send:

a. Your desired mutant
b. The parental wild-type strain
c. A characterized control mutant
d. Information on how the mutants were generated
e. A recommended check PCR protocol and primer sequences for performing check PCRs on the wild-type and control mutant

Conditions for sending out the mutants collaboratively:

a. We ask that 9 co-authors from our team be included on the first publication where the mutant we sent was used (even if this publication appears after we publish our manuscript). We feel that authorship is justified for generating and contributing a critical unpublished reagent. We realize that this is a lot of authors, but were unable to find a fair way to reduce this number, as all have contributed significantly to making the mutants. We will be happy to provide a methods section to describe the mutant generation. Please send a paper draft at least 2 weeks before submission to chlamymutants@gmail.com so that we have a chance to read the manuscript before submission.
b. You must agree to have the strain and its progeny distributed solely through the Chlamy Resource Center.

If you would like to receive additional mutants, or would rather not collaborate on these terms, the pilot collection of mutants will be made available with no authorship strings attached or limitations on number of mutants via the Chlamy Center upon the publication of our manuscript (target: early 2015).

Submit a mutant request by clicking this text (note- we will only send one mutant per PI, so be sure you choose carefully).