{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T08:59:11Z","timestamp":1790413151092,"version":"4.1.0"},"reference-count":49,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T00:00:00Z","timestamp":1784332800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T00:00:00Z","timestamp":1784332800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["IOS 2224861"],"award-info":[{"award-number":["IOS 2224861"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Plant Biotechnology Journal"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Our goal is to develop RNA\u2010guided engineering of the chloroplast genome using the CRISPR\/Cas9 system. We designed chloroplast minigenes to obtain properly sized single guide RNAs (sgRNAs) in tobacco chloroplasts. The sgRNA 5\u2032 end is defined by transcription from an rRNA operon promoter, and its 3\u2032 end by processing a downstream tRNA (trnG) or a hepatitis delta virus (HDV) ribozyme. Cas9 is expressed from a nuclear gene and is targeted to chloroplasts by fusion to a transit peptide. Cas9 incorporated the sgRNA and introduced double\u2010strand breaks in the plastid DNA (ptDNA). We report here that the double\u2010strand DNA break in the\n                    <jats:italic>ndhA<\/jats:italic>\n                    and\n                    <jats:italic>rpoC1<\/jats:italic>\n                    genes was repaired by microhomology\u2010mediated end joining (MMEJ), resulting in deletions in the ptDNA. We further showed that nuclear\u2010expressed sgRNA can be delivered into chloroplasts by fusion with a viroid RNA, as one possible approach for RNA\u2010guided engineering of the ptDNA without direct chloroplast genome transformation. These results are the first step of RNA\u2010guided editing of the chloroplast genome in any crop.\n                  <\/jats:p>","DOI":"10.1111\/pbi.70728","type":"journal-article","created":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T08:38:31Z","timestamp":1784363911000},"page":"5498-5509","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>RNA<\/scp>\n                    \u2010Guided Engineering of the Chloroplast Genome Enabled by Plastid\u2010Expressed Guide\n                    <scp>RNAs<\/scp>"],"prefix":"10.1111","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6274-7466","authenticated-orcid":false,"given":"Malihe","family":"Mirzaee","sequence":"first","affiliation":[{"name":"Waksman Institute of Microbiology, Rutgers University  Piscataway New Jersey 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