{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T09:01:13Z","timestamp":1768899673963,"version":"3.49.0"},"reference-count":42,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2024,9,20]],"date-time":"2024-09-20T00:00:00Z","timestamp":1726790400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2017YFD0200900"],"award-info":[{"award-number":["2017YFD0200900"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31101432"],"award-info":[{"award-number":["31101432"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Insect Science"],"published-print":{"date-parts":[[2025,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The clustered regularly interspaced small palindromic repeats (CRISPR) \/ CRISPR\u2010associated nuclease 9 (Cas9)\u2010mediated gene editing technology has revolutionized the study of fundamental biological questions in various insects. Diverse approaches have been developed to deliver the single\u2010guide RNA (sgRNA) and Cas9 to the nucleus of insect embryos or oocytes to achieve gene editing, including the predominant embryonic injection methods and alternative protocols through parental ovary delivery. However, a systematic comparative study of these approaches is limited, especially within a given insect. Here, we focused on revealing the detailed differences in CRISPR\/Cas9\u2010mediated gene editing between the embryo and ovary delivery methods in the beetle <jats:italic>Tribolium castaneum<\/jats:italic>, using the <jats:italic>cardinal<\/jats:italic> and <jats:italic>tyrosine hydroxylase<\/jats:italic> (<jats:italic>TH<\/jats:italic>) as reporter genes. We demonstrated that both genes could be efficiently edited by delivering Cas9\/sgRNA ribonucleoproteins to the embryos by microinjection, leading to the mutant phenotypes and indels in the target gene sites. Next, the Cas9\/sgRNA complex, coupled with a nanocarrier called Branched Amphiphilic Peptide Capsules (BAPC), were delivered to the ovaries of parental females to examine the efficacy of BAPC\u2010mediated gene editing. Although we observed that a small number of beetles\u2019 progeny targeting the <jats:italic>cardinal<\/jats:italic> exhibited the expected white\u2010eye phenotype, unexpectedly, no target DNA indels were found following subsequent sequencing analysis. In addition, we adopted a novel approach termed \u201cdirect parental\u201d CRISPR (DIPA\u2010CRISPR). However, we still failed to find gene\u2010editing events in the <jats:italic>cardinal<\/jats:italic> or <jats:italic>TH<\/jats:italic> gene\u2010targeted insects. Our results indicate that the conventional embryonic injection of CRISPR is an effective method to initiate genome editing in <jats:italic>T. castaneum<\/jats:italic>. However, it is inefficient by the parental ovary delivery approach.<\/jats:p>","DOI":"10.1111\/1744-7917.13447","type":"journal-article","created":{"date-parts":[[2024,9,20]],"date-time":"2024-09-20T03:29:28Z","timestamp":1726802968000},"page":"1185-1200","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Efficient CRISPR\u2010mediated genome editing can be initiated by embryonic injection but not by ovarian delivery in the beetle <i>Tribolium castaneum<\/i>"],"prefix":"10.1111","volume":"32","author":[{"given":"Zi\u2010Ling","family":"Liu","sequence":"first","affiliation":[{"name":"Key Laboratory of Plant Protection Resources and Pest Management of Ministry of Education, Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs College of Plant Protection, Northwest A&amp;F University  Yangling Shaanxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu\u2010Yu","family":"Zhou","sequence":"additional","affiliation":[{"name":"Key Laboratory of Plant Protection Resources and Pest Management of Ministry of Education, Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs College of Plant Protection, Northwest A&amp;F University  Yangling Shaanxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qiu\u2010Xuan","family":"Xu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Plant Protection Resources and Pest Management of Ministry of Education, Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs College of Plant Protection, Northwest A&amp;F University  Yangling Shaanxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xing\u2010Ce","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Plant Protection Resources and Pest Management of Ministry of Education, Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs College of Plant Protection, Northwest A&amp;F University  Yangling Shaanxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7686-0686","authenticated-orcid":false,"given":"Tong\u2010Xian","family":"Liu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Plant Protection Resources and Pest Management of Ministry of Education, Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs College of Plant Protection, Northwest A&amp;F University  Yangling Shaanxi China"},{"name":"Institute of Entomology Guizhou University  Guiyang Guizhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2205-0784","authenticated-orcid":false,"given":"Hong\u2010Gang","family":"Tian","sequence":"additional","affiliation":[{"name":"Key Laboratory of Plant Protection Resources and Pest Management of Ministry of Education, Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs College of Plant Protection, Northwest A&amp;F University  Yangling Shaanxi 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