{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T10:07:48Z","timestamp":1766138868867,"version":"build-2065373602"},"reference-count":22,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2024,9,6]],"date-time":"2024-09-06T00:00:00Z","timestamp":1725580800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"German Research Foundation","doi-asserted-by":"publisher","award":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"],"award-info":[{"award-number":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003390","name":"Fritz Thyssen Foundation","doi-asserted-by":"publisher","award":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"],"award-info":[{"award-number":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"]}],"id":[{"id":"10.13039\/501100003390","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Johannes and Frieda Marohn-Foundation","award":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"],"award-info":[{"award-number":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"]}]},{"name":"ELAN Fond of the University Hospital of Erlangen","award":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"],"award-info":[{"award-number":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"]}]},{"name":"Kastner-Stiftung","award":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"],"award-info":[{"award-number":["505539112\/KFO5024","GRK2599","TRR130","10.19.2.024MN","P117"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Introducing genetic material into hard-to-transfect mammalian cell lines and primary cells is often best achieved through retroviral infection. An ideal retroviral vector should offer a compact, selectable, and screenable marker while maximizing transgene delivery capacity. However, a previously published retroviral vector featuring an EGFP\/Puromycin fusion protein failed to meet these criteria in our experiments. We encountered issues such as low infection efficiency, weak EGFP fluorescence, and selection against infected cells. To address these shortcomings, we developed a novel retroviral vector based on the Moloney murine leukemia virus. This vector includes a compact bifunctional EGFP and Puromycin resistance cassette connected by a 2A peptide. Our extensively tested vector demonstrated superior EGFP expression, efficient Puromycin selection, and no growth penalty in infected cells compared with the earlier design. These benefits were consistent across multiple mammalian cell types, underscoring the versatility of our vector. In summary, our enhanced retroviral vector offers a robust solution for efficient infection, reliable detection, and effective selection in mammalian cells. Its improved performance and compact design make it an ideal choice for a wide range of applications involving precise genetic manipulation and characterization in cell-based studies.<\/jats:p>","DOI":"10.3390\/biom14091131","type":"journal-article","created":{"date-parts":[[2024,9,6]],"date-time":"2024-09-06T11:53:11Z","timestamp":1725623591000},"page":"1131","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["An Enhanced Retroviral Vector for Efficient Genetic Manipulation and Selection in Mammalian Cells"],"prefix":"10.3390","volume":"14","author":[{"given":"Jana","family":"Triller","sequence":"first","affiliation":[{"name":"Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger-Center of Molecular Medicine (NFZ), Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Gl\u00fcckstra\u00dfe 6, D-91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iryna","family":"Prots","sequence":"additional","affiliation":[{"name":"Department of Operative Dentistry and Periodontology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Gl\u00fcckstra\u00dfe 11, D-91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6332-8463","authenticated-orcid":false,"given":"Hans-Martin","family":"J\u00e4ck","sequence":"additional","affiliation":[{"name":"Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger-Center of Molecular Medicine (NFZ), Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Gl\u00fcckstra\u00dfe 6, D-91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0404-8186","authenticated-orcid":false,"given":"J\u00fcrgen","family":"Wittmann","sequence":"additional","affiliation":[{"name":"Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger-Center of Molecular Medicine (NFZ), Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg, Gl\u00fcckstra\u00dfe 6, D-91054 Erlangen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/1742-4690-7-8","article-title":"Hematopoietic stem cells and retroviral infection","volume":"7","author":"Banerjee","year":"2010","journal-title":"Retrovirology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1146\/annurev-immunol-051116-052155","article-title":"Immune Responses to Retroviruses","volume":"36","author":"Manel","year":"2018","journal-title":"Annu. 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