First Author | Mehlmann LM | Year | 2024 |
Journal | Genesis | Volume | 62 |
Issue | 5 | Pages | e23616 |
PubMed ID | 39487578 | Mgi Jnum | J:360169 |
Mgi Id | MGI:7787230 | Doi | 10.1002/dvg.23616 |
Citation | Mehlmann LM, et al. (2024) Generation and Characterization of a TRIM21 Overexpressing Mouse Line. Genesis 62(5):e23616 |
abstractText | Specific removal of a protein is a key to understanding its function. "Trim-Away" utilizes TRIM21, an antibody receptor and ubiquitin ligase, for acute and specific reduction of proteins. When TRIM21 is expressed in cells, introduction of a specific antibody causes rapid degradation of the targeted protein; however, TRIM21 is endogenously expressed in few cell types. We have generated a mouse line using CRISPR to insert a conditional overexpression cassette of TRIM21 into the safe harbor site, Rosa26. These conditionally-expressing mice can be bred to a wide variety of Cre mice to target cell-specific TRIM21 overexpression in different tissues. Zp3(Cre) mice expressed TRIM21 protein specifically in oocytes, whereas Hprt(Cre) mice expressed the protein globally. When TRIM21-overexpressing oocytes were microinjected with specific antibodies targeting either the IP(3) receptor or SNAP23, these proteins were effectively degraded. In addition, cortical neural cells from globally-overexpressing TRIM21 mice showed a dramatic reduction in IP(3) receptor protein within hours after electroporation of a specific antibody. These experiments confirm the effectiveness of the Trim-Away method for protein reduction. These mice should make a valuable addition to the broader research community, as a wide range of proteins and cell types can be studied using this method. |