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Title: Cross-Chiral, RNA-Catalyzed Exponential Amplification of RNA
Authors:
;
Award ID(s):
2114588
Publication Date:
NSF-PAR ID:
10334097
Journal Name:
Journal of the American Chemical Society
Volume:
143
Issue:
45
Page Range or eLocation-ID:
19160 to 19166
ISSN:
0002-7863
Sponsoring Org:
National Science Foundation
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  1. Abstract In vitro synthesized RNA is used widely in studies of RNA biology, biotechnology and RNA therapeutics. However, in vitro synthesized RNA often contains impurities, such as RNAs with lengths shorter and longer than the expected runoff RNA. We have recently confirmed that longer RNA products are formed predominantly via cis self-primed extension, in which released runoff RNA folds back on itself to prime its own RNA-templated extension. In the current work, we demonstrate that addition of a DNA oligonucleotide (capture DNA) that is complementary to the 3′ end of the expected runoff RNA effectively prevents self-primed extension, even under conditions commonly used for high RNA yields. Moreover, the presence of this competing capture DNA during ‘high yield’ transcription, leads to an increase in the yield of expected runoff RNA by suppressing the formation of undesired longer RNA byproducts.