The Fourth Chromosome Resource Project (PI: Stuart Newfeld, Arizona State University) is generating genetic reagents for genes located on the fourth chromosome of Drosophila melanogaster. The 'Associated Files' for this personal communication consist of .xlsx files that were submitted to FlyBase at the time that each set of fly stocks was generated and describe the details of the stocks.
The project is generating several classes of reagent, primarily:
1. Transgenic insertion stocks on the fourth chromosome containing a 'Double Header' (DH) trapping cassette. The DH cassette comprises two independent trapping modules oriented in opposite directions: 1. a protein-trap (PT) module for tagging endogenous proteins, composed of a splice acceptor site, a EGFP-FlAsH-StrepII-TEV-3xFlag ("GFSTF") multi-tag cassette and a splice donor site, and 2: a gene-trap (GT) module designed to truncate the endogenous gene into which it inserts, as it contains T2A-GAL4 followed by a polyadenylation signal. Depending on the orientation in which the DH cassette is inserted relative to the direction of transcription of the endogenous gene, the inserted element can either be in the 'protein-trap' or 'gene-trap' orientation for that gene.
See the following FlyBase dataset reports for details:
TI_set_Mi{DH}.Chr4 FBlc0003789
TI_set_TI{DH}.Chr4 FBlc0005341
In this case, the .xlsx files contain data primarily concerning the progenitor insertion line used, the phase of the donor construct used in the RMCE event, and the identities of any gene(s) in which the Double Header element is inserted.
The column "Trap? _Y_N_" for each gene indicates whether the cassette is (Y) or is not expected to trap (N) the gene.
2. A set of transgenic constructs for expression, under UAS control, of the coding sequence of fourth chromosome protein coding genes.
See the following FlyBase dataset report for details:
FCRP_UAS_fly_cDNA FBlc0003790
3. A set of transgenic constructs for expression, under UAS control, of the coding sequence of human orthologs of fourth chromosome protein coding genes.
FCRP_UAS_human_cDNA FBlc0003844
4. CRISPR/Cas9-generated mutations of fourth chromosome protein coding genes in which the mutation has been fully sequenced.