{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T15:24:33Z","timestamp":1777908273106,"version":"3.51.4"},"reference-count":39,"publisher":"SAGE Publications","issue":"6","license":[{"start":{"date-parts":[[2024,12,1]],"date-time":"2024-12-01T00:00:00Z","timestamp":1733011200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["The CRISPR Journal"],"published-print":{"date-parts":[[2024,12,1]]},"abstract":"<jats:sec>\n                    <jats:title>Background:<\/jats:title>\n                    <jats:p>\n                      Melanoma remains one of the most challenging cancers to treat effectively with drug resistant remaining a constant concern, primarily with activating\n                      <jats:italic toggle=\"yes\">BRAF<\/jats:italic>\n                      mutations. Mutations in the\n                      <jats:italic toggle=\"yes\">BRAF<\/jats:italic>\n                      gene appear in approximately 50% of patients, 90% of which are V600E. Two frontline\n                      <jats:italic toggle=\"yes\">BRAF<\/jats:italic>\n                      inhibitors (BRAFi), vemurafenib and dabrafenib, are frequently used to treat unresectable or metastatic\n                      <jats:italic toggle=\"yes\">BRAF<\/jats:italic>\n                      V600E melanoma. Initial response rates are high, but soon thereafter, 70\u201380% of patients develop resistance to treatment within a year. A major mechanism of resistance is the generation of a secondary Q61K mutation in the\n                      <jats:italic toggle=\"yes\">NRAS<\/jats:italic>\n                      gene.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods:<\/jats:title>\n                    <jats:p>\n                      We have developed an approach in which a CRISPR-Cas complex can be designed to distinguish between mutant genes enabling resistance to standard care in tumor cells and normal genomes of healthy cells. For the first time, we demonstrated the utility of two CRISPR-directed mutation-specific editing approaches to restore BRAFi sensitivity in\n                      <jats:italic toggle=\"yes\">BRAF<\/jats:italic>\n                      <jats:sup>V600E<\/jats:sup>\n                      \/\n                      <jats:italic toggle=\"yes\">NRAS<\/jats:italic>\n                      <jats:sup>Q61K<\/jats:sup>\n                      resistant A375 cells.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      We utilize an AsCas12a protospacer adjacent motif site created by the\n                      <jats:italic toggle=\"yes\">NRAS<\/jats:italic>\n                      Q61K mutation and the Q61K mutation in the critical seed region of an SaCas9 sgRNA for Q61K-selective targeting. We show here that both approaches allow for effective\n                      <jats:italic toggle=\"yes\">NRAS<\/jats:italic>\n                      targeting of only mutated-Q61K and after CRISPR-directed Q61K-targeting, previously resistant A375 cells are re-sensitized to BRAFi treatment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion:<\/jats:title>\n                    <jats:p>Our data support the feasibility of the development of CRISPR-Cas therapeutic approaches to the treatment of melanoma. Successful therapeutic CRISPR-directed gene editing would enable both specific and efficient editing of a mutation-specific targeting approach eliminate concern for on- and off-target damage to the genomes of healthy cells.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1089\/crispr.2024.0003","type":"journal-article","created":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T07:45:03Z","timestamp":1728546303000},"page":"366-373","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["Mutation-Specific CRISPR Targeting with SaCas9 and AsCas12a Restores Therapeutic Sensitivity in Treatment-Resistant Melanoma"],"prefix":"10.1177","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5178-6953","authenticated-orcid":false,"given":"Brett M.","family":"Sansbury","sequence":"first","affiliation":[{"name":"Gene Editing Institute, ChristianaCare Health System, Newark, Delaware."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sophia B.","family":"Masciarelli","sequence":"additional","affiliation":[{"name":"Gene Editing Institute, ChristianaCare Health System, Newark, Delaware."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Salma","family":"Kaouser","sequence":"additional","affiliation":[{"name":"Gene Editing Institute, ChristianaCare Health System, Newark, Delaware."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olivia M.","family":"Tharp","sequence":"additional","affiliation":[{"name":"Gene Editing Institute, ChristianaCare Health System, Newark, Delaware."},{"name":"Department of Medical and Molecular Sciences, University of Delaware, Newark, Delaware."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kelly H.","family":"Banas","sequence":"additional","affiliation":[{"name":"Gene Editing Institute, ChristianaCare Health System, Newark, Delaware."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4680-5325","authenticated-orcid":false,"given":"Eric B.","family":"Kmiec","sequence":"additional","affiliation":[{"name":"Gene Editing Institute, ChristianaCare Health System, Newark, Delaware."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2024,12,12]]},"reference":[{"key":"e_1_3_6_2_2","unstructured":"Melanoma Skin Cancer Statistics | American Cancer Society. n.d. 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