Highly multiplexed, image-based pooled screens in primary cells and tissues with PerturbView
Highly multiplexed, image-based pooled screens in primary cells and tissues with PerturbView
Optical pooled screening (OPS) is a highly scalable method for linking image-based phenotypes with cellular perturbations. However, it has thus far been restricted to relatively low-plex phenotypic readouts in cancer cell lines in culture, due to limitations associated with in situ sequencing (ISS) of perturbation barcodes. Here, we developed PerturbView, an OPS technology that leverages in vitro transcription (IVT) to amplify barcodes prior to ISS, enabling screens with highly multiplexed phenotypic readouts across diverse systems, including primary cells and tissues. We demonstrate PerturbView in iPSC-derived neurons, primary immune cells, and tumor tissue sections from animal models. In a screen of immune signaling pathways in primary bone marrow-derived macrophages, PerturbView uncovered both known and novel regulators of NFΚB signaling. Furthermore, we combined PerturbView with spatial transcriptomics in tissue sections from a mouse xenograft model, paving the way to in vivo screens with rich optical and transcriptomic phenotypes. PerturbView broadens the scope of OPS to a wide range of models and applications.
Lu Cherry Sakura、Lubeck Eric、Wu Ping、Hu Xiaoyu、Foo Conrad、Town Jason P、Wei Runmin、Celen Cemre、Kljavin Noelyn、Kayagaki Nobuhiko、Garraway Levi、Melo Felipe de Sousa e、Chen Yushu、Bravo Hector Corrada、Li Bo、McGinnis Lisa、Kudo Takamasa、Jesudason Rajiv、Rozenblatt-Rosen Orit、Hleap Jose Sergio、Heinlein Melanie、Moncada Reuben、Rios Antonio、Hockendorf Burkhard、Regev Aviv、Jiang Jian、Kornfeld Opher、Meireles Ana M、Singh Avtar、Chandrasekar Vineethkrishna、Cai Shuangyi、Gould Joshua
生物科学研究方法、生物科学研究技术基础医学分子生物学
Lu Cherry Sakura,Lubeck Eric,Wu Ping,Hu Xiaoyu,Foo Conrad,Town Jason P,Wei Runmin,Celen Cemre,Kljavin Noelyn,Kayagaki Nobuhiko,Garraway Levi,Melo Felipe de Sousa e,Chen Yushu,Bravo Hector Corrada,Li Bo,McGinnis Lisa,Kudo Takamasa,Jesudason Rajiv,Rozenblatt-Rosen Orit,Hleap Jose Sergio,Heinlein Melanie,Moncada Reuben,Rios Antonio,Hockendorf Burkhard,Regev Aviv,Jiang Jian,Kornfeld Opher,Meireles Ana M,Singh Avtar,Chandrasekar Vineethkrishna,Cai Shuangyi,Gould Joshua.Highly multiplexed, image-based pooled screens in primary cells and tissues with PerturbView[EB/OL].(2025-03-28)[2025-04-26].https://www.biorxiv.org/content/10.1101/2023.12.26.573143.点此复制
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