Pipeline Status Explanations
The following list defines the different statuses that EUCOMM report on the constructs in their pipeline.
Mice - Genotype confirmed (M-GC)
The genotype of testcross offspring has been confirmed molecularly.
Mice - Germline transmission (M-GLT)
Test crosses of chimaeras indicate germline transmission of the ES cell mutation based on coat color.
Mice - Microinjection in progress (M-MIP)
Targeted ES cells have been microinjected for the purpose of creating mutant mice for distribution.
ES Cells - Targeting Confirmed (ES-TC)
Correctly targeted ES cell clones have passed LR-PCR/sequenced-based QC. These ES cell clones can be requested from the EUCOMM repositories.
ES Cells - Targeting Unsuccessful - Project Terminated (ES-PT)
Electroporation of the final targeting construct containing a promoter-driven selectable cassette yielded an insufficient number of colonies for analysis. Work on this particular gene will cease and the gene may be passed to Regeneron or Norcomm.
ES Cells - No QC Positives (ES-NQP)
ES Cells - Electroporation Unsuccessful (ES-EU)
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.
ES Cells - Electroporation in Progress (ES-EP)
Electroporation of the final targeting construct is underway.
Vector - DNA Not Suitable for Electroporation (ES-DNA)
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 repositories.
Vector Complete (TVC)
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 repositories.
Vector - Initial Attempt Unsuccessful (TV-IAU)
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 Unsuccessful - Alternate Design in Progress (TV-ADP)
The construction of the targeting vector was not successful or did not pass sequence-based QC. Recombineering will be repeated using an alternative valid conditional design (allele).
ES Cells - Conditional Gene Trap (ESC-GT)
DNA sequence from the gene trap clones is used to identify the intronic insertion site, sequence from the targeting vectors is used to define the precise allele structures and BAC ends are used to position the BACs in the genome. This information is mapped onto the mouse genome sequence using standard methods.