{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T16:01:48Z","timestamp":1790438508135,"version":"4.1.0"},"reference-count":15,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Single\u2010cell RNA sequencing (scRNA\u2010seq) reveals the transcriptional heterogeneity of cells, revolutionizing our understanding of cellular processes. However, the static snapshots obtained from scRNA\u2010seq fail to reveal the time\u2010resolved dynamics of transcription, which impedes critical insights into various biological processes, such as cellular differentiation, embryonic development, disease progression, and responses to external stimuli. Here, we describe Well\u2010TEMP\u2010seq, a protocol for massively parallel profiling of the temporal dynamics of single\u2010cell gene expression. Well\u2010TEMP\u2010seq combines metabolic RNA labeling with a microwell\u2010based scRNA\u2010seq method, Well\u2010paired\u2010seq, to distinguish newly transcribed RNAs marked by T\u2010to\u2010C substitutions from pre\u2010existing RNAs in each of thousands of single cells. Well\u2010TEMP\u2010seq is high\u2010throughput, cost\u2010effective, accurate, and provides a low cell loss rate and high single cell\/bead pairing efficiency. More importantly, Well\u2010TEMP\u2010seq can be easily set up in other labs, and the loading of cells and beads can be easily accomplished by an optical microscope and a pipette. We believe that Well\u2010TEMP\u2010seq will be widely adopted and help researchers perform transformative research to unveil the dynamics of single\u2010cell gene expression in diverse biological processes. \u00a9 2026 Wiley Periodicals LLC.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Well\u2010paired\u2010seq chip fabrication\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Well\u2010TEMP\u2010seq sample processing\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Bioinformatics analysis\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70458","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T16:50:12Z","timestamp":1788195012000},"source":"Crossref","is-referenced-by-count":0,"title":["Massively Parallel Profiling of Single\u2010Cell RNA Dynamics Using Well\u2010TEMP\u2010seq"],"prefix":"10.1002","volume":"6","author":[{"given":"Di","family":"Wang","sequence":"first","affiliation":[{"name":"MOE Key Laboratory of Spectrochemical Analysis &amp; Instrumentation, State Key Laboratory of Vaccines for Infectious Diseases, Xiang An Biomedicine Laboratory, Fujian Key Laboratory of Chemical Biology, Department of Chemical Biology, College of Chemistry and Chemical Engineering Xiamen University  Xiamen China"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiqi","family":"Lv","sequence":"additional","affiliation":[{"name":"School of Life Sciences Xiamen University  Xiamen China"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shichao","family":"Lin","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Spectrochemical Analysis &amp; Instrumentation, State Key Laboratory of Vaccines for Infectious Diseases, Xiang An Biomedicine Laboratory, Fujian Key Laboratory of Chemical Biology, Department of Chemical Biology, College of Chemistry and Chemical Engineering Xiamen University  Xiamen China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aax3072"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587\u2010020\u20100480\u20109"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/smtd.202100111"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg3055"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature06758"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010019\u20101369\u2010y"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43586\u2010022\u201000157\u2010z"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1534\u20105807(03)00373\u20103"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010019\u201011028\u20109"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1039\/D1LC00607J"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010023\u201036902\u20105"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010020\u20100935\u20104"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature00927"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.017400"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/smtd.202200341"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70458","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpz1.70458","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70458","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T15:55:18Z","timestamp":1790438118000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70458"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,31]]},"references-count":15,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1002\/cpz1.70458"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70458","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,31]]},"article-number":"e70458"}}