{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T13:49:51Z","timestamp":1790430591674,"version":"4.1.0"},"reference-count":31,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2023,5,25]],"date-time":"2023-05-25T00:00:00Z","timestamp":1684972800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,6,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>The degradation tag (dTAG) system for target protein degradation can remove proteins from biological systems without the drawbacks of some genetic methods, such as slow kinetics, lack of reversibility, low specificity, and the inability to titrate dosage. These drawbacks can make it difficult to compare toxicity resulting from genetic and pharmacological interventions, especially in vivo. Because the dTAG system has not been studied extensively in vivo, we explored the use of this system to study the physiological sequalae resulting from CDK2 or CDK5 degradation in adult mice. Mice with homozygous knock-in of the dTAG sequence onto CDK2 and CDK5 were born at Mendelian ratios despite decreased CDK2 or CDK5 protein levels in comparison with wild-type mice. In bone marrow cells and duodenum organoids derived from these mice, treatment with the dTAG degrader dTAG-13 resulted in rapid and robust protein degradation but caused no appreciable change in viability or the transcriptome. Repeated delivery of dTAG-13 in vivo for toxicity studies proved challenging; we explored multiple formulations in an effort to maximize degradation while minimizing formulation-related toxicity. Degradation of CDK2 or CDK5 in all organs except the brain, where dTAG-13 likely did not cross the blood brain barrier, only caused microscopic changes in the testis of CDK2dTAG mice. These findings were corroborated with conditional CDK2 knockout in adult mice. Our results suggest that the dTAG system can provide robust protein degradation in vivo and that loss of CDK2 or CDK5 in adult mice causes no previously unknown phenotypes.<\/jats:p>","DOI":"10.1093\/toxsci\/kfad049","type":"journal-article","created":{"date-parts":[[2023,5,25]],"date-time":"2023-05-25T19:35:34Z","timestamp":1685043334000},"page":"53-69","source":"Crossref","is-referenced-by-count":21,"title":["Use of the dTAG system <i>in vivo<\/i> to degrade CDK2 and CDK5 in adult mice and explore potential safety liabilities"],"prefix":"10.1093","volume":"194","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3912-0571","authenticated-orcid":false,"given":"Paul","family":"Yenerall","sequence":"first","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae","family":"Sung","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiran","family":"Palyada","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessie","family":"Qian","sequence":"additional","affiliation":[{"name":"Pfizer , Groton, Connecticut 06340, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3988-1947","authenticated-orcid":false,"given":"Seda","family":"Arat","sequence":"additional","affiliation":[{"name":"Pfizer , Groton, Connecticut 06340, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven W","family":"Kumpf","sequence":"additional","affiliation":[{"name":"Pfizer , Groton, Connecticut 06340, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shih-Wen","family":"Wang","sequence":"additional","affiliation":[{"name":"Pfizer , Cambridge, Massachusetts 02139, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen","family":"Biddle","sequence":"additional","affiliation":[{"name":"Pfizer , Groton, Connecticut 06340, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos","family":"Esparza","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanie","family":"Chang","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wesley","family":"Scott","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Walter","family":"Collette","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taylor-Symon","family":"Winrow","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tim","family":"Affolter","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0730-8836","authenticated-orcid":false,"given":"Norimitsu","family":"Shirai","sequence":"additional","affiliation":[{"name":"Pfizer , Groton, Connecticut 06340, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephane","family":"Thibault","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6640-0718","authenticated-orcid":false,"given":"Julia","family":"Wang","sequence":"additional","affiliation":[{"name":"Pfizer , Cambridge, Massachusetts 02139, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ling","family":"Liu","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary","family":"Bauchmann","sequence":"additional","affiliation":[{"name":"Pfizer , Groton, Connecticut 06340, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4298-1973","authenticated-orcid":false,"given":"Jessica","family":"Frey","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefanus","family":"Steyn","sequence":"additional","affiliation":[{"name":"Pfizer , Cambridge, Massachusetts 02139, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aida","family":"Sacaan","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allison","family":"Vitsky","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5988-7388","authenticated-orcid":false,"given":"Youngwook","family":"Ahn","sequence":"additional","affiliation":[{"name":"Pfizer , Cambridge, Massachusetts 02139, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tom","family":"Paul","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6119-0253","authenticated-orcid":false,"given":"Lawrence","family":"Lum","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5479-3400","authenticated-orcid":false,"given":"Jon","family":"Oyer","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy","family":"Yang","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4662-3727","authenticated-orcid":false,"given":"Wenyue","family":"Hu","sequence":"additional","affiliation":[{"name":"Pfizer , San Diego, California 92121, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,5,25]]},"reference":[{"key":"2023062820430803100_kfad049-B1","doi-asserted-by":"crossref","first-page":"1068","DOI":"10.1016\/j.devcel.2022.03.013","article-title":"Rapid and efficient degradation of endogenous proteins in vivo identifies stage-specific roles of RNA pol II pausing in mammalian development","volume":"57","author":"Abuhashem","year":"2022","journal-title":"Dev. Cell"},{"key":"2023062820430803100_kfad049-B2","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1111\/vcp.12773","article-title":"Interpretative considerations for clinical pathology findings in nonclinical toxicology studies","volume":"48","author":"Aulbach","year":"2019","journal-title":"Vet. Clin. Pathol"},{"key":"2023062820430803100_kfad049-B3","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1038\/nm.1754","article-title":"Chemical control of protein stability and function in living mice","volume":"14","author":"Banaszynski","year":"2008","journal-title":"Nat. Med"},{"key":"2023062820430803100_kfad049-B4","doi-asserted-by":"crossref","first-page":"1775","DOI":"10.1016\/j.cub.2003.09.024","article-title":"CDK2 knockout mice are viable","volume":"13","author":"Berthet","year":"2003","journal-title":"Curr. Biol"},{"key":"2023062820430803100_kfad049-B5","doi-asserted-by":"crossref","first-page":"5495","DOI":"10.1038\/s41467-022-33246-4","article-title":"Systematic profiling of conditional degron tag technologies for target validation studies","volume":"13","author":"Bondeson","year":"2022","journal-title":"Nat. Commun"},{"key":"2023062820430803100_kfad049-B6","doi-asserted-by":"crossref","first-page":"1831","DOI":"10.1021\/acschembio.5b00442","article-title":"Haloprotacs: use of small molecule protacs to induce degradation of halotag fusion proteins","volume":"10","author":"Buckley","year":"2015","journal-title":"ACS Chem. Biol"},{"key":"2023062820430803100_kfad049-B7","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1038\/nchembio.1869","article-title":"Tunable and reversible drug control of protein production via a self-excising degron","volume":"11","author":"Chung","year":"2015","journal-title":"Nat. Chem. Biol"},{"key":"2023062820430803100_kfad049-B8","doi-asserted-by":"crossref","first-page":"e0152989","DOI":"10.1371\/journal.pone.0152989","article-title":"In vivo performance and properties of tamoxifen metabolites for creert2 control","volume":"11","author":"Felker","year":"2016","journal-title":"PLoS One"},{"key":"2023062820430803100_kfad049-B9","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1126\/science.8016642","article-title":"Deletion of a DNA polymerase beta gene segment in t cells using cell type-specific gene targeting","volume":"265","author":"Gu","year":"1994","journal-title":"Science"},{"key":"2023062820430803100_kfad049-B10","first-page":"626","article-title":"The ten most common adverse drug reactions (ADRs) in oncology patients: do they matter to you?","volume":"12","author":"Lau","year":"2004","journal-title":"Support. Care Cancer"},{"key":"2023062820430803100_kfad049-B11","doi-asserted-by":"crossref","first-page":"7274","DOI":"10.1038\/ncomms8274","article-title":"CDK5 controls lymphatic vessel development and function by phosphorylation of Foxc2","volume":"6","author":"Liebl","year":"2015","journal-title":"Nat. Commun"},{"key":"2023062820430803100_kfad049-B12","doi-asserted-by":"crossref","first-page":"2464","DOI":"10.3390\/cells10092464","article-title":"The atypical cyclin-dependent kinase 5 (CDK5) guards podocytes from apoptosis in glomerular disease while being dispensable for podocyte development","volume":"10","author":"Mangold","year":"2021","journal-title":"Cells"},{"key":"2023062820430803100_kfad049-B13","doi-asserted-by":"crossref","first-page":"6088","DOI":"10.18632\/oncotarget.6842","article-title":"Inhibition of endothelial CDK5 reduces tumor growth by promoting non-productive angiogenesis","volume":"7","author":"Merk","year":"2016","journal-title":"Oncotarget"},{"key":"2023062820430803100_kfad049-B14","doi-asserted-by":"crossref","first-page":"4687","DOI":"10.1038\/s41467-020-18377-w","article-title":"Rapid and direct control of target protein levels with VHL-recruiting dTAG molecules","volume":"11","author":"Nabet","year":"2020","journal-title":"Nat. Commun"},{"key":"2023062820430803100_kfad049-B15","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1038\/s41589-018-0021-8","article-title":"The dTAG system for immediate and target-specific protein degradation","volume":"14","author":"Nabet","year":"2018","journal-title":"Nat. Chem. Biol"},{"key":"2023062820430803100_kfad049-B16","doi-asserted-by":"crossref","first-page":"R53","DOI":"10.1186\/bcr3154","article-title":"Functional characterization of the 19Q12 amplicon in grade III breast cancers","volume":"14","author":"Natrajan","year":"2012","journal-title":"Breast Cancer Res"},{"key":"2023062820430803100_kfad049-B17","doi-asserted-by":"crossref","first-page":"11173","DOI":"10.1073\/pnas.93.20.11173","article-title":"Targeted disruption of the cyclin-dependent kinase 5 gene results in abnormal corticogenesis, neuronal pathology and perinatal death","volume":"93","author":"Ohshima","year":"1996","journal-title":"Proc. Natl. Acad. Sci. U.S.A"},{"key":"2023062820430803100_kfad049-B18","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1038\/ng1232","article-title":"Cyclin-dependent kinase 2 is essential for meiosis but not for mitotic cell division in mice","volume":"35","author":"Ortega","year":"2003","journal-title":"Nat. Genet"},{"key":"2023062820430803100_kfad049-B19","doi-asserted-by":"crossref","first-page":"3566","DOI":"10.3390\/cancers12123566","article-title":"Combined CDK2 and CDK4\/6 inhibition overcomes palbociclib resistance in breast cancer by enhancing senescence","volume":"12","author":"Pandey","year":"2020","journal-title":"Cancers"},{"key":"2023062820430803100_kfad049-B20","doi-asserted-by":"crossref","first-page":"2627","DOI":"10.1002\/jps.24506","article-title":"Plasma protein binding of challenging compounds","volume":"104","author":"Riccardi","year":"2015","journal-title":"J. Pharm. Sci"},{"key":"2023062820430803100_kfad049-B21","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1038\/ng1117","article-title":"A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference","volume":"33","author":"Rubinson","year":"2003","journal-title":"Nat. Genet"},{"key":"2023062820430803100_kfad049-B22","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1038\/nature06046","article-title":"CDK1 is sufficient to drive the mammalian cell cycle","volume":"448","author":"Santamar\u00eda","year":"2007","journal-title":"Nature"},{"key":"2023062820430803100_kfad049-B23","doi-asserted-by":"crossref","first-page":"190287","DOI":"10.1098\/rsob.190287","article-title":"A kinase of many talents: non-neuronal functions of CDK5 in development and disease","volume":"10","author":"Sharma","year":"2020","journal-title":"Open Biol"},{"key":"2023062820430803100_kfad049-B24","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1146\/annurev-cellbio-092910-154023","article-title":"Cyclin-dependent kinases in brain development and disease","volume":"27","author":"Su","year":"2011","journal-title":"Annu. Rev. Cell Dev. Biol"},{"key":"2023062820430803100_kfad049-B25","doi-asserted-by":"crossref","first-page":"13865","DOI":"10.1002\/anie.202004087","article-title":"Development of CDK2 and CDK5 dual degrader TMX-2172","volume":"59","author":"Teng","year":"2020","journal-title":"Angew. Chem. Int. Ed"},{"key":"2023062820430803100_kfad049-B26","doi-asserted-by":"crossref","first-page":"1169","DOI":"10.1200\/JCO.18.00925","article-title":"Cyclin e1 expression and palbociclib efficacy in previously treated hormone receptor-positive metastatic breast cancer","volume":"37","author":"Turner","year":"2019","journal-title":"J. Clin. Oncol"},{"key":"2023062820430803100_kfad049-B27","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1016\/j.cell.2013.04.025","article-title":"One-step generation of mice carrying mutations in multiple genes by CRISPR\/Cas-mediated genome engineering","volume":"153","author":"Wang","year":"2013","journal-title":"Cell"},{"key":"2023062820430803100_kfad049-B28","doi-asserted-by":"crossref","first-page":"2743","DOI":"10.1038\/s41467-020-16559-0","article-title":"Quantifying CDK inhibitor selectivity in live cells","volume":"11","author":"Wells","year":"2020","journal-title":"Nat. Commun"},{"key":"2023062820430803100_kfad049-B29","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1016\/j.cell.2013.08.022","article-title":"One-step generation of mice carrying reporter and conditional alleles by CRISPR\/Cas-mediated genome engineering","volume":"154","author":"Yang","year":"2013","journal-title":"Cell"},{"key":"2023062820430803100_kfad049-B30","doi-asserted-by":"crossref","first-page":"20801","DOI":"10.18632\/oncotarget.4600","article-title":"Cyclin-dependent kinase 2 is an ideal target for ovary tumors with elevated cyclin E1 expression","volume":"6","author":"Yang","year":"2015","journal-title":"Oncotarget"},{"key":"2023062820430803100_kfad049-B31","doi-asserted-by":"crossref","first-page":"5701","DOI":"10.1038\/s41467-020-19532-z","article-title":"The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice","volume":"11","author":"Yesbolatova","year":"2020","journal-title":"Nat. Commun"}],"container-title":["Toxicological Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/toxsci\/advance-article-pdf\/doi\/10.1093\/toxsci\/kfad049\/50515333\/kfad049.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/toxsci\/article-pdf\/194\/1\/53\/50734994\/kfad049.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/toxsci\/article-pdf\/194\/1\/53\/50734994\/kfad049.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,6,28]],"date-time":"2023-06-28T22:16:22Z","timestamp":1687990582000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/toxsci\/article\/194\/1\/53\/7179808"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,5,25]]},"references-count":31,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,5,25]]},"published-print":{"date-parts":[[2023,6,28]]}},"URL":"https:\/\/doi.org\/10.1093\/toxsci\/kfad049","relation":{},"ISSN":["1096-6080","1096-0929"],"issn-type":[{"value":"1096-6080","type":"print"},{"value":"1096-0929","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2023,7,1]]},"published":{"date-parts":[[2023,5,25]]}}}