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Impurities arising from incomplete synthesis, degradation, or side reactions can compromise pharmacological performance and induce off-target or immune responses. For long, structured oligonucleotides such as CRISPR guides, purification can be costly in both resources and yield. To overcome these limitations, we developed a purification method based on a 5\u2032-terminal dioctyloxytrityl (DOT) protecting group. In this purification method, DOT-protected full-length compounds show high-resolution separation from truncated impurities. The DOT group can then be removed under mild, rapid, and scalable conditions. DOT chemistry is analogous to \u201cDMT-ON\u201d approaches that are well-known and widely used in the field, and is fully compatible with existing synthesis workflows, but dramatically improves HPLC purification relative to DMT-ON approaches by providing higher retention and thus better resolution to enable the purification of long, complex, and chemically modified sequences.<\/jats:p>","DOI":"10.1021\/acs.joc.6c00856","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T09:02:03Z","timestamp":1786438923000},"page":"11286-11292","source":"Crossref","is-referenced-by-count":0,"title":["Improved Purification\nof sgRNA and pegRNA by Use of\na 5\u2032-Terminal Dioctyloxytrityl (DOT) Protecting Group"],"prefix":"10.1021","volume":"91","author":[{"given":"Atish A.","family":"Wagh","sequence":"first","affiliation":[{"name":"University of Massachusetts Chan Medical School , , , ,","place":["Worcester, Massachusetts, United States, 01605"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5706-1734","authenticated-orcid":true,"given":"Jonathan K.","family":"Watts","sequence":"additional","affiliation":[{"name":"University of Massachusetts Chan Medical School , , , ,","place":["Worcester, Massachusetts, United States, 01605"]},{"name":"University of Massachusetts Chan Medical School , , , ,","place":["Worcester, Massachusetts, United States, 01605"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"key":"2026082211140902500_cit1","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1038\/d41573-020-00078-0","article-title":"RNA therapeutics on the rise","volume":"19","author":"Wang","year":"2020","journal-title":"Nat. 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