Genome-Wide High-Throughput RNAi Screening for Identification of Genes Involved in Protein Production.

Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE

Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,

RNA Interference* ; High-Throughput Screening Assays*/methods ; RNA, Small Interfering*/genetics ; Transfection*/methods; Humans ; Animals ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Cell Survival/genetics

With an increasing number of blockbuster drugs being recombinant mammalian proteins, protein production platforms that focus on mammalian proteins have had a profound impact in many areas of basic and applied research. Many groups, both academic and industrial, have been focusing on developing cost-effective methods to improve the production of mammalian proteins that would support potential therapeutic applications. As it stands, while a wide range of platforms have been successfully developed for laboratory use, the majority of biologicals are still produced in mammalian cell lines due to the requirement for posttranslational modification and the biosynthetic complexity of target proteins. An unbiased high-throughput RNAi screening approach can be an efficient tool to identify target genes involved in recombinant protein production. Here, we describe the process of optimizing the transfection conditions, performing the genome-wide siRNA screen, the activity and cell viability assays, and the validation transfection to identify genes involved with protein expression.
(© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

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Keywords: HEK 293; Protein production; Screen; siRNA

0 (RNA, Small Interfering)
0 (Recombinant Proteins)

Date Created: 20240626 Date Completed: 20240626 Latest Revision: 20240626

20240627

10.1007/978-1-0716-3878-1_20

38926288