{"id":137451,"date":"2022-03-28T20:03:25","date_gmt":"2022-03-29T03:03:25","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2022\/03\/crispr-cas9-mediated-genome-editing-as-a-therapeutic-approach-for-leber-congenital-amaurosis-10"},"modified":"2022-03-28T20:03:25","modified_gmt":"2022-03-29T03:03:25","slug":"crispr-cas9-mediated-genome-editing-as-a-therapeutic-approach-for-leber-congenital-amaurosis-10","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2022\/03\/crispr-cas9-mediated-genome-editing-as-a-therapeutic-approach-for-leber-congenital-amaurosis-10","title":{"rendered":"CRISPR\/Cas9-Mediated Genome Editing as a Therapeutic Approach for Leber Congenital Amaurosis 10"},"content":{"rendered":"<p><a class=\"aligncenter blog-photo\" href=\"https:\/\/lifeboat.com\/blog.images\/crispr-cas9-mediated-genome-editing-as-a-therapeutic-approach-for-leber-congenital-amaurosis-10.jpg\"><\/a><\/p>\n<p>Circa 2017 \ud83d\ude00<\/p>\n<hr>\n<p>As the most common subtype of Leber congenital amaurosis (LCA), LCA10 is a severe retinal dystrophy caused by mutations in the <i>CEP290<\/i> gene. The most frequent mutation found in patients with LCA10 is a deep intronic mutation in <i>CEP290<\/i> that generates a cryptic splice donor site. The large size of the <i>CEP290<\/i> gene prevents its use in adeno-associated virus (AAV)-mediated gene augmentation therapy. Here, we show that targeted genomic deletion using the clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas9 system represents a promising therapeutic approach for the treatment of patients with LCA10 bearing the <i>CEP290<\/i> splice mutation. We generated a cellular model of LCA10 by introducing the <i>CEP290<\/i> splice mutation into 293FT cells and we showed that guide RNA pairs coupled with SpCas9 were highly efficient at removing the intronic splice mutation and restoring the expression of wild-type <i>CEP290<\/i>. In addition, we demonstrated that a dual AAV system could effectively delete an intronic fragment of the <i>Cep290<\/i> gene in the mouse retina. To minimize the immune response to prolonged expression of SpCas9, we developed a self-limiting CRISPR\/Cas9 system that minimizes the duration of SpCas9 expression. These results support further studies to determine the therapeutic potential of CRISPR\/Cas9-based strategies for the treatment of patients with LCA10.<\/p>\n<p><strong class=\"\"> Keywords: <\/strong> <i>CEP290<\/i>; CRISPR\/Cas9; LCA10.<\/p>\n<p>Copyright \u00a9 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Circa 2017 \ud83d\ude00 As the most common subtype of Leber congenital amaurosis (LCA), LCA10 is a severe retinal dystrophy caused by mutations in the CEP290 gene. The most frequent mutation found in patients with LCA10 is a deep intronic mutation in CEP290 that generates a cryptic splice donor site. The large size of the CEP290 [\u2026]<\/p>\n","protected":false},"author":513,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[11,412],"tags":[],"class_list":["post-137451","post","type-post","status-publish","format-standard","hentry","category-biotech-medical","category-genetics"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/137451","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/users\/513"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=137451"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/137451\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=137451"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=137451"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=137451"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}