Tg(tetO-Ctsb/RB1)14Smrs

A 1583 bp segement of the human RB1 cDNA (bases 1243-2826, amino-acids 369-896) was cloned into the pCS2+CB-Myc6 vector. This vector contains a 1012 bp (337 amino-acids) long construct of bp 35-1047 of Ctsb with 6x Myc tags fused to its C-terminus. The resulting fusion product is an 865 aa peptide comprising Ctsb, six Myc tags and Rb1. The fusion gene was then cloned into a pTet-splice vector which brings the fusion gene under the control of the tetO promoter and terminates the gene with an SV40 intron and polyA signals. This transgenic construct was used to generate several founder mice, of which this line was used for further study. The transgene results in a tetracycline-inducibele dominant negative allele that disrupts the function of Rb1-containing multimeric complexes by shuttling them to the lysosome for degradation. (J:242452)