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We have used long single\u2010stranded DNAs having phosphorothioate modifications as donor templates for HDR at targeted genomic sites using the Clustered Regularly Interspaced Short Palindromic Repeats\/CRISPR\u2010associated protein 9 (CRISPR\/Cas9) system. First, <jats:italic>X. tropicalis tyr<\/jats:italic> mutant (i.e., albino) embryos were successfully rescued: partially pigmented tadpoles were seen in up to 35% of injected embryos, demonstrating the potential for efficient insertion of targeted point mutations. Second, in order to demonstrate the ability to tag genes with fluorescent proteins (FPs), we targeted the melanocyte\u2010specific gene <jats:italic>slc45a2.L<\/jats:italic> of <jats:italic>X. laevis<\/jats:italic> to label it with the <jats:italic>Superfolder green<\/jats:italic> FP (<jats:italic>sfGFP<\/jats:italic>), seeing mosaic expression of sfGFP in melanophores in up to 20% of injected tadpoles. Tadpoles generated by these two approaches were raised to sexual maturity, and shown to successfully transmit HDR constructs through the germline with precise targeting and seamless recombination. F1 embryos showed rescue of the <jats:italic>tyr<\/jats:italic> mutation (<jats:italic>X. tropicalis<\/jats:italic>) and tagging in the appropriate pigment cell\u2010specific manner of <jats:italic>slc45a2.L<\/jats:italic> with <jats:italic>sfGFP<\/jats:italic> (<jats:italic>X. laevis<\/jats:italic>).<\/jats:p>","DOI":"10.1002\/dvg.23366","type":"journal-article","created":{"date-parts":[[2020,4,11]],"date-time":"2020-04-11T13:35:22Z","timestamp":1586612122000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Simple embryo injection of long single\u2010stranded donor templates with the <scp>CRISPR<\/scp>\/Cas9 system leads to homology\u2010directed repair in <scp><i>Xenopus tropicalis<\/i><\/scp> and <scp><i>Xenopus laevis<\/i><\/scp>"],"prefix":"10.1002","volume":"58","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5531-6710","authenticated-orcid":false,"given":"Takuya","family":"Nakayama","sequence":"first","affiliation":[{"name":"Department of Biology University of Virginia  Charlottesville Virginia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4539-5280","authenticated-orcid":false,"given":"Robert M.","family":"Grainger","sequence":"additional","affiliation":[{"name":"Department of Biology University of Virginia  Charlottesville Virginia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5171-5685","authenticated-orcid":false,"given":"Sang\u2010Wook","family":"Cha","sequence":"additional","affiliation":[{"name":"School of Natural Sciences University of Central Missouri  Warrensburg Missouri"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,4,11]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-019-0444-0"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.152967"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.22719"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0925-4773(02)00219-8"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12915-018-0530-7"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.117.300468"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1163\/002829680X00177"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)00200-2"},{"issue":"1","key":"e_1_2_7_10_1","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1242\/dev.99.1.15","article-title":"Distribution, expression and germ line transmission of exogenous DNA sequences following microinjection into Xenopus laevis eggs","volume":"99","author":"Etkin L. 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