{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T05:06:57Z","timestamp":1790744817855,"version":"4.1.0"},"reference-count":50,"publisher":"SAGE Publications","issue":"4","license":[{"start":{"date-parts":[[2022,8,1]],"date-time":"2022-08-01T00:00:00Z","timestamp":1659312000000},"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":[[2022,8,1]]},"abstract":"<jats:p>\n                    Epidermolysis bullosa simplex (EBS) is a rare mechanobullous disease caused by dominant-negative mutations in either keratin 5 (\n                    <jats:italic toggle=\"yes\">KRT5<\/jats:italic>\n                    ) or keratin 14 (\n                    <jats:italic toggle=\"yes\">KRT14<\/jats:italic>\n                    ) genes. Until now, there is no cure for EBS and the care is primarily palliative. The discovery of the clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 system raised hope for the treatment of EBS and many other autosomal dominant diseases by mutant allele-specific gene disruption. In this study, we aim to disrupt the mutant allele for the heterozygous EBS pathogenic variation c.449T&gt;C (p.Leu150Pro) within\n                    <jats:italic toggle=\"yes\">KRT5<\/jats:italic>\n                    . This mutation generates, naturally, a novel protospacer-adjacent motif for the endonuclease\n                    <jats:italic toggle=\"yes\">Streptococcus pyogenes Cas9<\/jats:italic>\n                    . Thus, we designed a single-guide RNA that guides the Cas9 to introduce a DNA cleavage of the mutant allele in patient's keratinocytes. Then, transfected cells were single-cell cloned and analyzed by deep sequencing. The expression of\n                    <jats:italic toggle=\"yes\">KRT5<\/jats:italic>\n                    and\n                    <jats:italic toggle=\"yes\">KRT14<\/jats:italic>\n                    was quantified, and the keratin intermediate filament stability was assessed. Results showed successful stringent mutant allele-specific knockout. An absence of synthesis of mutant transcript was further confirmed indicating permanent mutant allele-specific inactivation. Edited EBS patient keratinocytes produced a lower amount of K5 and K14 proteins compared with nonedited EBS cells, and no disturbance of cellular properties was observed.\n                  <\/jats:p>","DOI":"10.1089\/crispr.2021.0132","type":"journal-article","created":{"date-parts":[[2022,7,21]],"date-time":"2022-07-21T11:29:50Z","timestamp":1658402990000},"page":"586-597","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":13,"title":["Allele-Specific Inactivation of an Autosomal Dominant Epidermolysis Bullosa Simplex Mutation Using CRISPR-Cas9"],"prefix":"10.1177","volume":"5","author":[{"given":"Mbarka","family":"Bchetnia","sequence":"first","affiliation":[{"name":"D\u00e9partement des Sciences Fondamentales and Universit\u00e9 Laval, Qu\u00e9bec, Canada."},{"name":"Centre intersectoriel en sant\u00e9 durable (CISD), Universit\u00e9 du Qu\u00e9bec \u00e0 Chicoutimi (UQAC), Saguenay, Canada; Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca","family":"Dionne Gagn\u00e9","sequence":"additional","affiliation":[{"name":"D\u00e9partement des Sciences Fondamentales and Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie","family":"Powell","sequence":"additional","affiliation":[{"name":"Service de Dermatologie, CHU Sainte-Justine, Montr\u00e9al, Canada; Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles","family":"Morin","sequence":"additional","affiliation":[{"name":"Centre Int\u00e9gr\u00e9 Universitaire de Sant\u00e9 et de Services Sociaux du Saguenay-Lac-Saint-Jean, H\u00f4pital Universitaire de Chicoutimi, Saguenay, Canada; and Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine","family":"McCuaig","sequence":"additional","affiliation":[{"name":"Service de Dermatologie, CHU Sainte-Justine, Montr\u00e9al, Canada; Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrey","family":"Dup\u00e9r\u00e9e","sequence":"additional","affiliation":[{"name":"Centre Int\u00e9gr\u00e9 Universitaire de Sant\u00e9 et de Services Sociaux du Saguenay-Lac-Saint-Jean, H\u00f4pital Universitaire de Chicoutimi, Saguenay, Canada; and Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucie","family":"Germain","sequence":"additional","affiliation":[{"name":"D\u00e9partement de chirurgie, Facult\u00e9 de m\u00e9decine, Centre de Recherche en Organog\u00e9n\u00e8se Exp\u00e9rimentale de l'Universit\u00e9 Laval\/LOEX, and Universit\u00e9 Laval, Qu\u00e9bec, Canada."},{"name":"Centre de recherche du CHU de Qu\u00e9bec, Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacques P.","family":"Tremblay","sequence":"additional","affiliation":[{"name":"Centre de recherche du CHU de Qu\u00e9bec, Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5526-9945","authenticated-orcid":false,"given":"Catherine","family":"Laprise","sequence":"additional","affiliation":[{"name":"D\u00e9partement des Sciences Fondamentales and Universit\u00e9 Laval, Qu\u00e9bec, Canada."},{"name":"Centre intersectoriel en sant\u00e9 durable (CISD), Universit\u00e9 du Qu\u00e9bec \u00e0 Chicoutimi (UQAC), Saguenay, Canada; Universit\u00e9 Laval, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2022,8,12]]},"reference":[{"key":"e_1_3_6_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.det.2009.10.003"},{"key":"e_1_3_6_3_2","first-page":"637","article-title":"Mutations in KRT5 and KRT14 cause epidermolysis bullosa simplex in 75% of the patients","volume":"164","author":"Bolling MC","year":"2011","unstructured":"Bolling MC, , Lemmink HH, , Jansen GH, et al. Mutations in KRT5 and KRT14 cause epidermolysis bullosa simplex in 75% of the patients. Br J Dermatol. 2011; 164:637\u2013644. DOI: 10.1111\/j.1365\u20132133.2010.10146.x","journal-title":"Br J Dermatol"},{"key":"e_1_3_6_4_2","volume-title":"In: GeneReviews\u00ae\n                  . (Adam MP, Ardinger HH, Pagon RA, et al.; eds)","author":"Pfendner EG","year":"1993","unstructured":"Pfendner EG, , Bruckner AL. Epidermolysis bullosa simplex. In: GeneReviews\u00ae. (Adam MP, Ardinger HH, Pagon RA, et al.; eds). Seattle, WA: University of Washington, Seattle. 1993."},{"key":"e_1_3_6_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaad.2014.01.903"},{"key":"e_1_3_6_6_2","first-page":"119","article-title":"Revised clinical and laboratory criteria for subtypes of inherited epidermolysis bullosa","volume":"24","author":"Fine JD","year":"1991","unstructured":"Fine JD, , Bauer EA, , Briggaman RA, et al. Revised clinical and laboratory criteria for subtypes of inherited epidermolysis bullosa. A consensus report by the Subcommittee on Diagnosis and Classification of the National Epidermolysis Bullosa Registry. J Am Acad Dermatol., 1991; 24:119\u2013135. DOI: 10.1016\/0190-9622(91)70021-s","journal-title":"A consensus report by the Subcommittee on Diagnosis and Classification of the National Epidermolysis Bullosa Registry. 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