Nucleoside analogs in ADAR guide strands targeting 5'-UA̲ sites.
Nucleoside analogs in ADAR guide strands targeting 5'-UA̲ sites.
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靶向5 '-UA位点的阿达尔引导链中的核苷类似物。
DOI：
10.1039/d2cb00165a
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发表时间：
2023-01-04
影响因子：
4.1
通讯作者：
Beal, Peter A.
中科院分区：
文献类型：
--
作者：
Brinkman, Hannah F.;Jauregui Matos, Victorio;Mendoza, Herra G.;Doherty, Erin E.;Beal, Peter A.
Adenosine deaminases that act on RNA (ADARs) can be directed to predetermined sites in transcriptomes by forming duplex structures with exogenously delivered guide RNAs (gRNAs). They can then catalyze the hydrolytic deamination of adenosine to inosine in double stranded RNA, which is read as guanosine during translation. High resolution structures of ADAR2-RNA complexes revealed a unique conformation for the nucleotide in the guide strand base paired to the editing site's 5′ nearest neighbor (−1 position). Here we describe the effect of 16 different nucleoside analogs at this position in a gRNA that targets a 5′-UA̲-3′ site. We found that several analogs increase editing efficiency for both catalytically active human ADARs. In particular, 2′-deoxynebularine (dN) increased the ADAR1 and ADAR2 in vitro deamination rates when at the −1 position of gRNAs targeting the human MECP2 W104X site, the mouse IDUA W392X site, and a site in the 3′-UTR of human ACTB. Furthermore, a locked nucleic acid (LNA) modification at the −1 position was found to eliminate editing. When placed −1 to a bystander editing site in the MECP2 W104X sequence, bystander editing was eliminated while maintaining on-target editing. In vitro trends for four −1 nucleoside analogs were validated by directed editing of the MECP2 W104X site expressed on a reporter transcript in human cells. This work demonstrates the importance of the −1 position of the gRNA to ADAR editing and discloses nucleoside analogs for this site that modulate ADAR editing efficiency. Adenosine deaminases acting on RNAs (ADARs) can be directed to correct RNA mutations with complementary guide strands. We introduce nucleoside analogs at the −1 position of the guide RNA to enhance target editing and decrease off-target editing.
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影响因子：
15
作者：
Doherty EE;Wilcox XE;van Sint Fiet L;Kemmel C;Turunen JJ;Klein B;Tantillo DJ;Fisher AJ;Beal PA
通讯作者：
Beal PA
影响因子：
14.9
作者：
Thuy-Boun AS;Thomas JM;Grajo HL;Palumbo CM;Park S;Nguyen LT;Fisher AJ;Beal PA
通讯作者：
Beal PA
影响因子：
14.9
作者：
Fritzell, Kajsa;Xu, Li-Di;Ohman, Marie
通讯作者：
Ohman, Marie
影响因子：
46.9
作者：
Monian, Prashant;Shivalila, Chikdu;Vargeese, Chandra
通讯作者：
Vargeese, Chandra
影响因子：
4.6
作者：
Fukuda M;Umeno H;Nose K;Nishitarumizu A;Noguchi R;Nakagawa H
通讯作者：
Nakagawa H