{"id":110275,"date":"2020-07-21T10:22:46","date_gmt":"2020-07-21T17:22:46","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2020\/07\/crispr-c-to-g-base-editors-for-inducing-targeted-dna-transversions-in-human-cells"},"modified":"2020-07-21T10:22:46","modified_gmt":"2020-07-21T17:22:46","slug":"crispr-c-to-g-base-editors-for-inducing-targeted-dna-transversions-in-human-cells","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2020\/07\/crispr-c-to-g-base-editors-for-inducing-targeted-dna-transversions-in-human-cells","title":{"rendered":"CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells"},"content":{"rendered":"

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

CRISPR-guided DNA cytosine and adenine base editors are widely used for many applications1<\/a>,2<\/a>,3<\/a>,4<\/a><\/sup> but primarily create DNA base transitions (that is, pyrimidine-to-pyrimidine or purine-to-purine). Here we describe the engineering of two base editor architectures that can efficiently induce targeted C-to-G base transversions, with reduced levels of unwanted C-to-W (W = A or T) and indel mutations. One of these C-to-G base editors (CGBE1), consists of an RNA-guided Cas9 nickase, an Escherichia coli <\/i>\u2013derived uracil DNA N-glycosylase (eUNG) and a rat APOBEC1 cytidine deaminase variant (R33A) previously shown to have reduced off-target RNA and DNA editing activities5<\/a>,6<\/a><\/sup>. We show that CGBE1 can efficiently induce C-to-G edits, particularly in AT-rich sequence contexts in human cells. We also removed the eUNG domain to yield miniCGBE1, which reduced indel frequencies but only modestly decreased editing efficiency. CGBE1 and miniCGBE1 enable C-to-G edits and will serve as a basis for optimizing C-to-G base editors for research and therapeutic applications.<\/p>\n","protected":false},"excerpt":{"rendered":"

CRISPR-guided DNA cytosine and adenine base editors are widely used for many applications1,2,3,4 but primarily create DNA base transitions (that is, pyrimidine-to-pyrimidine or purine-to-purine). Here we describe the engineering of two base editor architectures that can efficiently induce targeted C-to-G base transversions, with reduced levels of unwanted C-to-W (W = A or T) and indel [\u2026]<\/p>\n","protected":false},"author":396,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[11,38],"tags":[],"class_list":["post-110275","post","type-post","status-publish","format-standard","hentry","category-biotech-medical","category-engineering"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/110275","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\/396"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=110275"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/110275\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=110275"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=110275"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=110275"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}