{"id":11149,"date":"2018-12-14T10:50:50","date_gmt":"2018-12-14T09:50:50","guid":{"rendered":"http:\/\/www.wjst.de\/blog\/?p=11149"},"modified":"2019-06-17T10:47:31","modified_gmt":"2019-06-17T09:47:31","slug":"off-target-sites-of-the-crisp-cas-babies","status":"publish","type":"post","link":"https:\/\/www.wjst.de\/blog\/sciencesurf\/2018\/12\/off-target-sites-of-the-crisp-cas-babies\/","title":{"rendered":"Off target sites of the CRISPR CAS babies"},"content":{"rendered":"

Unfortunately we don’t know the genome of the Chinese parents which would be necessary for any prediction of genome mutations in the twins . So far we only know one confirmed off target site from Dr. He’s Hongkong slides that reports 5 mismatches.<\/p>\n

\"reported<\/a>
reported off target site<\/figcaption><\/figure>\n

At least I would expect for statistical reasons, some 1 bp mismatch, some 2bp, some 3bp…, but not just one 5bp off target site.
\nSo we could\u00a0check several<\/a> online services with the know sgRNA sequence that Dr. He used.<\/p>\n

CRISPOR<\/a><\/span><\/p>\n

\"\"<\/a><\/p>\n

CRISPOR finds one region with 2 mismatches (intergenic) and 13 with 3 mismatches (7 intergenic hits, 5x\u00a0introns (ST8SIA6, CTD-2532D12.5, CNTN5, C2D3, RABGAP1) and 1x exon of ACACA).\u00a0This doesn’t match the reported off target sequence.<\/p>\n

Cas-OFFinder<\/a><\/span><\/p>\n

\"\"<\/a><\/p>\n

Cas-OFFinder identifies 5 regions with 2 mismatches (1x intergenic, 2x sites in LIMCH1, 1x intergenic, 1 in\u00a0BPIFC).<\/p>\n

CCtop<\/a>\u00a0(yes, I was waiting for that name :-)<\/span><\/p>\n

\"\"<\/a><\/p>\n

COS finds only one intergenic region with 3 mismatches.<\/p>\n

CRISPRSCAN<\/a><\/span><\/p>\n

\"\"<\/a><\/p>\n

has the best explanation of what it does but cannot search from sgRNA to off-target as many other online tools.<\/p>\n

OFFspotter<\/a><\/p>\n

\"\"<\/a><\/p>\n

is spotting 1 intergenic region with 2 mismatches, \u00a03 with 3 mismatches (2 intergenic, 1 in\u00a0RP11-238K6.1).<\/p>\n

Taken these five different predictions together, I can’t make conclusion but think that we need to resequence the families with much higher coverage (150-200x fold coverage) and even compare the mutations with the phenotype of the children.<\/p>\n\n

 <\/p>\n

\n CC-BY-NC Science Surf , accessed 05.04.2026<\/span>\n <\/div>","protected":false},"excerpt":{"rendered":"

Unfortunately we don’t know the genome of the Chinese parents which would be necessary for any prediction of genome mutations in the twins . So far we only know one confirmed off target site from Dr. He’s Hongkong slides that reports 5 mismatches. At least I would expect for statistical reasons, some 1 bp mismatch, … Continue reading Off target sites of the CRISPR CAS babies<\/span> →<\/span><\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[2],"tags":[3079],"class_list":["post-11149","post","type-post","status-publish","format-standard","hentry","category-genetics-biology","tag-genome-editing"],"_links":{"self":[{"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/posts\/11149","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/comments?post=11149"}],"version-history":[{"count":10,"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/posts\/11149\/revisions"}],"predecessor-version":[{"id":12716,"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/posts\/11149\/revisions\/12716"}],"wp:attachment":[{"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/media?parent=11149"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/categories?post=11149"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.wjst.de\/blog\/wp-json\/wp\/v2\/tags?post=11149"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}