{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T19:45:30Z","timestamp":1792611930819,"version":"4.5.0"},"reference-count":85,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T00:00:00Z","timestamp":1792540800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T00:00:00Z","timestamp":1792540800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"publisher","award":["BB\/W014610\/1"],"award-info":[{"award-number":["BB\/W014610\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Yeast"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Yeasts are model organisms and protein production hosts of choice, in part due to their ease of genetic manipulation. Yeast transformation with DNA on plasmids or integrated into the genome has been developed since the 1970s and is a routine method in many laboratories. However, transformation frequencies are surprisingly low, with common methods yielding around 1% of transformed cells in budding yeast\n                    <jats:italic>Saccharomyces cerevisiae<\/jats:italic>\n                    . In alternative species, there is lower transformation frequency and unreasonably large quantities of input DNA may be required. Surprisingly, the mechanisms by which yeast cells take up DNA are not known. Here we argue that three approaches could deliver a step\u2010change in yeast transformation frequency and efficiency. First, investigating the mechanisms of DNA uptake. The requirement for salts and extraneous polymers\u2014carrier DNA and PEG\u2014in standard protocols suggests that phase separation of DNA may enhance DNA uptake into cells or subsequent nuclear delivery. Second, accounting for the physiological or growth state of the yeast including cell cycle phase and nutrient or stress conditions. Third, systematically determining the factors that affect transformation efficiency using statistical Design of Experiments. These approaches could allow superefficient transformation of\n                    <jats:italic>S. cerevisiae<\/jats:italic>\n                    , and accelerate genetic manipulation of non\u2010model yeasts.\n                  <\/jats:p>","DOI":"10.1002\/yea.70051","type":"journal-article","created":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T19:22:31Z","timestamp":1792610551000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Approaches to Superefficient Yeast Transformations: DNA Phase Separation, Cell Growth State and Systematic Experimental Design"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4429-6232","authenticated-orcid":false,"given":"Alistair","family":"Elfick","sequence":"first","affiliation":[{"name":"Centre for Engineering Biology University of Edinburgh Edinburgh UK"},{"name":"School of Engineering University of Edinburgh Edinburgh UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8025-6361","authenticated-orcid":false,"given":"Edward W. J.","family":"Wallace","sequence":"additional","affiliation":[{"name":"Centre for Engineering Biology University of Edinburgh Edinburgh UK"},{"name":"School of Biological Sciences University of Edinburgh Edinburgh UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,21]]},"reference":[{"issue":"8","key":"e_1_2_9_2_1","doi-asserted-by":"crossref","first-page":"1964","DOI":"10.1021\/acs.jced.2c00042","article-title":"Nucleic Acid Partitioning in PEG\u2010Ficoll Protocells","volume":"67","author":"Ahmed T.","year":"2022","journal-title":"Journal of Chemical & Engineering Data"},{"issue":"9","key":"e_1_2_9_3_1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0163049","article-title":"Chemical Inhibitors of Non\u2010Homologous End Joining Increase Targeted Construct Integration in Cryptococcus Neoformans","volume":"11","author":"Arras S. D. 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