- The targeting strategy allows the generation of a conditional Knock-Out (KO) and a constitutive KO of the Brd4 gene.
- The targeting strategy is based on the NCBI transcript NM_020508.3.
- Exon 3 contains the translation initiation codon.
- Exons 6 and 7 have been flanked by loxP sites (size of loxP-flanked region is 1.8 kb).
- The positive selection marker (Puromycin resistance - PuroR) has been flanked by FRT sites and inserted into intron 5.
- The targeting vector has been generated using BAC clones from the C57BL/6J RPCIB-731 BAC library and will be transfected into the TaconicArtemis C57BL/6N Tac ES cell line.
- Homologous recombinant clones will be isolated using positive (PuroR) and negative (Thymidine kinase - Tk) selections.
- The conditional KO allele is obtained after Flp-mediated removal of the selection marker.
- The constitutive KO allele is obtained after Cre-mediated recombination. Deletion of exons 6 and 7 should result in the loss of function of the Brd4 gene by generating a frameshift from exon 5 to exons 8-11 (premature Stop codon in exon 8). In addition the resulting transcript may be a target for Non-sense Mediated RNA Decay and may therefore not be expressed at significant level.
- The remaining recombination sites will be located in non-conserved regions of the genome.
Gene Family: Transcriptional regulator
Genetic Background: C57BL/6
Gene Symbol: Brd4
Gene Accession Number: ENSMUST00000003726
Allele Type: cKO
Official Gene Name: bromodomain containing 4