International Knockout Mouse Consortium - Knowledgebase

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How can I determine the knockout status of my gene?

(AUG4UAG)

Enter the gene symbol into the search box on the upper right and search. The resulting query summary lists one record (row) for each gene that matches your query.

The Gene column displays the gene symbol, chromosome, and location.

The Knockout Project Status column displays the current status of the gene in the following knockout project pipelines: KOMP-CSD, KOMP-Regeneron, EUCOMM, NorCOMM. The “Details” link leads to a page that provides more information about knockout attempts for the gene.

The Other Resources column displays information about mutant ES cells and mice for the gene that are available through the International Mouse Strain Resource (IMSR) or the International Gene Trap Consortium (IGTC). Also listed are targeted mutations and other mutations recorded in MGI.

The gene status details page (reached by following the “Details” link from the results summary) includes status information about all knockout attempts for the selected gene. If knockout attempts have lead to the generation of a targeting vector, or to more advanced stages, links to graphics detailing targeting vector and allele designs are provided. If IKMC products are available, order links to the corresponding repositories are provided.

The Regeneron pipeline comprises the following statuses:

  • Regeneron Selected
  • Parental BAC Obtained
  • Design Finished/Oligos Ordered
  • Targeting Vector QC Completed
  • Vector Electroporated into ES Cells
  • ES cell colonies picked
  • ES cell colonies screened / QC no positives
  • ES cell colonies screened / QC one positive
  • ES cell colonies screened / QC positives
  • ES Cell Clone Microinjected
  • Germline Transmission Achieved

The EUCOMM and CSD pipelines comprise the following statuses:

Mice

  • Genotype confirmed: The genotype of testcross germline offspring has been confirmed molecularly.
  • Germline transmission: Test crosses of chimaeras indicate germline transmission of the ES cell mutation based on coat color.
  • Microinjection in progress: Targeted ES cells have been microinjected for the purpose of creating mutant mice for distribution.

ES Cells

  • Targeting Confirmed: Correctly targeted ES cell clones have passed LR-PCR/sequenced-based QC. These ES cell clones can be requested from the EUCOMM/KOMP repositories.
  • Targeting Unsuccessful Project Terminated: No targeted clones have been obtained and the project has been terminated.
  • No QC Positives: ES cell clones were picked and screened, but did not pass LR-PCR/sequenced-based QC. These ES cell clones may be re-screened with additional genotyping primers.
  • Electroporation Unsuccessful: Electroporation performed, but yielded few or no colonies
  • Electroporation in Progress: Electroporation of the final targeting construct is underway.

Vector

  • Vector Complete: A final targeting vector (post Gateway) has been constructed, has passed sequence-based QC, and is available for use in ES cells. These vectors can be requested from the EUCOMM/KOMP repositories.
  • DNA Not Suitable for Electroporation: A final targeting vector (post Gateway) has been constructed, has passed sequence-based QC, but has failed gel-based QC prior to electroporation (low DNA yield, incomplete digestion or multiple DNA bands). These vectors will be re-prepped for electroporation and can be requested from the EUCOMM/KOMP repositories.
  • Vector Complete Project Terminated: A project which has targeting vector but after re-synthesis still no successful ES cell, so it terminated. Also vectors constructed but not optimal for targeting the gene of interest
  • Vector Unsuccessful - Project Terminated: A project is terminated when two attempts at producing a valid conditional design (allele) have failed.
  • Vector Unsuccessful Alternate Design in Progress: The vector for a failed targeting project is in the process of being re-designed
  • Vector Initial Attempt Unsuccessful: The construction of a final targeting construct, following the Gateway exchange reaction, was not successful or did not pass sequence-based QC; pending a second attempt.
  • Vector Construction in Progress: Oligos for recombineering have been ordered for the construction of the targeting vectors with vector construction to follow
  • Design Completed: The gene has been manually annotated by the Havana group and the design of a conditional allele has been generated.
  • Alternate Design Requested: The gene’s earlier designs have not succeeded, and a new design is needed.

Mutant ES cells or mice for your gene generated by other projects (i.e. outside the IKMC effort) might already be publicly available from the International Gene Trap Consortium (IGTC) or through the International Mouse Strain Resource (IMSR). Please check the Other Resources column for this information.

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