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Making And Using Antibodies: A Practical Handbook, Second Edition

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Other CRISPR systems, specifically the Type VI CRISPR enzymes Cas13a/C2c2 and Cas13b, target RNA rather than DNA. Fusing a hyperactive adenosine deaminase that acts on RNA, ADAR2(E488Q), to catalytically dead Cas13b creates a programmable RNA base editor that converts adenosine to inosine in RNA (termed REPAIR). Since inosine is functionally equivalent to guanosine, the result is an A->G change in RNA. The catalytically inactive Cas13b ortholog from Prevotella sp., dPspCas13b, does not appear to require a specific sequence adjacent to the RNA target, making this a very flexible editing system. Editors based on a second ADAR variant, ADAR2(E488Q/T375G), display improved specificity, and editors carrying the delta-984-1090 ADAR truncation retain RNA editing capabilities and are small enough to be packaged in AAV particles.




Making and Using Antibodies: A Practical Handbook, Second Edition


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