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However, since typical chemical routes towards activating RNA substrates are incompatible with ribozyme catalysis, it remains unclear how prebiotic systems generated and sustained pools of activated building blocks needed to form increasingly larger and complex RNA. Herein, we demonstrate in situ activation of RNA substrates under reaction conditions amenable to catalysis by the hairpin ribozyme. We found that diamidophosphate (DAP) and imidazole drive the formation of 2\u2032,3\u2032\u2010cyclic phosphate RNA mono\u2010 and oligonucleotides from monophosphorylated precursors in frozen water\u2010ice. This long\u2010lived activation enables iterative enzymatic assembly of long RNAs. Our results provide a plausible scenario for the generation of higher\u2010energy substrates required to fuel ribozyme\u2010catalyzed RNA synthesis in the absence of a highly evolved metabolism.<\/jats:p>","DOI":"10.1002\/anie.202010918","type":"journal-article","created":{"date-parts":[[2020,10,30]],"date-time":"2020-10-30T23:03:40Z","timestamp":1604099020000},"page":"2952-2957","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Prebiotically Plausible RNA Activation Compatible with Ribozyme\u2010Catalyzed Ligation"],"prefix":"10.1002","volume":"60","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9959-9994","authenticated-orcid":false,"given":"Emilie Yeonwha","family":"Song","sequence":"first","affiliation":[{"name":"Max Planck Institute of Biochemistry  Am Klopferspitz 18 82152 Martinsried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2249-8116","authenticated-orcid":false,"given":"Eddy Ivanhoe","family":"Jim\u00e9nez","sequence":"additional","affiliation":[{"name":"Department of Chemistry The Scripps Research Institute  10550 North Torrey Pines Road La Jolla CA 92037 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huacan","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Chemistry The Scripps Research Institute  10550 North Torrey Pines Road La Jolla CA 92037 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2455-8706","authenticated-orcid":false,"given":"Kristian","family":"Le Vay","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Biochemistry  Am Klopferspitz 18 82152 Martinsried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5238-610X","authenticated-orcid":false,"given":"Ramanarayanan","family":"Krishnamurthy","sequence":"additional","affiliation":[{"name":"Department of Chemistry The Scripps Research Institute  10550 North Torrey Pines Road La Jolla CA 92037 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8005-1657","authenticated-orcid":false,"given":"Hannes","family":"Mutschler","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Biochemistry  Am Klopferspitz 18 82152 Martinsried Germany"},{"name":"Technical University Dortmund  Otto-Hahn-Strasse 4a 44227 Dortmund Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,12,10]]},"reference":[{"key":"e_1_2_2_1_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201704048"},{"key":"e_1_2_2_1_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201704048"},{"key":"e_1_2_2_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.7690155"},{"key":"e_1_2_2_3_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1060786"},{"key":"e_1_2_2_4_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1610103113"},{"key":"e_1_2_2_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1405"},{"key":"e_1_2_2_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chemgeo.2017.11.008"},{"key":"e_1_2_2_7_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1006683507864"},{"key":"e_1_2_2_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01751807"},{"key":"e_1_2_2_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja00845a050"},{"key":"e_1_2_2_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(82)90169-3"},{"key":"e_1_2_2_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1914367117"},{"key":"e_1_2_2_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.2878"},{"key":"e_1_2_2_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201903808"},{"key":"e_1_2_2_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201903808"},{"key":"e_1_2_2_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)41278-7"},{"key":"e_1_2_2_15_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1327.1998.2570286.x"},{"key":"e_1_2_2_16_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1999.2543"},{"key":"e_1_2_2_17_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkh601"},{"key":"e_1_2_2_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja4127714"},{"key":"e_1_2_2_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.2251"},{"key":"e_1_2_2_20_1","doi-asserted-by":"publisher","DOI":"10.3390\/life4020131"},{"key":"e_1_2_2_21_1","doi-asserted-by":"publisher","DOI":"10.5194\/bg-3-243-2006"},{"key":"e_1_2_2_22_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.43022"},{"key":"e_1_2_2_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms1076"},{"key":"e_1_2_2_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbdv.200890141"},{"key":"e_1_2_2_25_1","first-page":"2789","volume":"26","author":"Walba H.","year":"1961","journal-title":"J. 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