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Poly(ADP-ribose) polymerase inhibitors (PARPi) have shown remarkable efficacy, especially in BRCA-mutant patients, and are approved as maintenance therapy to prevent recurrence after initial response to chemotherapy. However, the development of PARPi resistance poses a major clinical challenge. This study utilized a whole-genome CRISPR-Cas9 genetic screening to identify genes associated with PARPi sensitivity upon knockout. Based on the screening and validated through further experiments, we confirmed that CLK1 knockdown is synthetically lethal with PARPi in ovarian cancer. The combination of the PARPi Olaparib and CLK1 inhibitor TG003 exhibited potent anti-proliferative effects both in vitro and in vivo. Mechanistically, CLK1 inhibition downregulated the functional ERCC1-202 isoform, resulting in enhanced DNA damage and apoptosis. Our findings reveal a novel mechanism underlying PARPi sensitivity and suggest that targeting CLK1 in combination with PARPi may represent a promising therapeutic strategy for PARPi-resistant ovarian cancer.<\/jats:p>","DOI":"10.1093\/procel\/pwaf091","type":"journal-article","created":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T12:47:18Z","timestamp":1762087638000},"page":"248-262","source":"Crossref","is-referenced-by-count":2,"title":["Genome-wide CRISPR-Cas9 screening identifies CLK1 inhibition as a strategy to restore PARP inhibitor sensitivity via ERCC1 isoform switching"],"prefix":"10.1093","volume":"17","author":[{"given":"Chaohua","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]},{"name":"Cancer Institute, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yutuan","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Gynecological Oncology, Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences , Hangzhou 310024,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiana","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengdong","family":"Ni","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyu","family":"Xia","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lihua","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 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200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuqi","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meiqin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiajia","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohua","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 200032,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center , Shanghai 200032,","place":["China"]},{"name":"Department of Oncology, Shanghai Medical College, Fudan University , Shanghai 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