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Direct capture of CRISPR guides enables scalable, multiplexed, and multi-omic Perturb-seq

Joseph M. Replogle, Albert Xu, Thomas M. Norman, Elliott J. Meer, Jessica M. Terry, Daniel Riordan, Niranjan Srinivas, Tarjei S. Mikkelsen, Jonathan S. Weissman, Britt Adamson
doi: https://doi.org/10.1101/503367
Joseph M. Replogle
Tetrad Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA 94158, USA Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158, USA
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Albert Xu
Medical Scientist Training Program, University of California, San Francisco, San Francisco, CA 94158, USA Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158, USA
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Thomas M. Norman
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158, USA
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Elliott J. Meer
10x Genomics Inc., Pleasanton, California, 94566, USA
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Jessica M. Terry
10x Genomics Inc., Pleasanton, California, 94566, USA
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Daniel Riordan
10x Genomics Inc., Pleasanton, California, 94566, USA
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Niranjan Srinivas
10x Genomics Inc., Pleasanton, California, 94566, USA
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Tarjei S. Mikkelsen
10x Genomics Inc., Pleasanton, California, 94566, USA
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Jonathan S. Weissman
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158, USA
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  • For correspondence: jonathan.weissman@ucsf.edu badamson@princeton.edu
Britt Adamson
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158, USA Present address: Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544, USA Present address: Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA
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  • For correspondence: jonathan.weissman@ucsf.edu badamson@princeton.edu
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Abstract

Pairing CRISPR-based genetic screens with single-cell transcriptional phenotypes (Perturb-seq) has advanced efforts to explore the function of mammalian genes and genetic networks. We present strategies for Perturb-seq that enable direct capture of CRISPR sgRNAs within 3’ or 5’ single-cell RNA-sequencing libraries using the 10x Genomics platform. This technology greatly expands the accessibility, scalability, and flexibility of Perturb-seq, specifically enabling use with programmed combinatorial perturbations and multiplexing with multi-omic measurements.

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Posted December 21, 2018.
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Direct capture of CRISPR guides enables scalable, multiplexed, and multi-omic Perturb-seq
Joseph M. Replogle, Albert Xu, Thomas M. Norman, Elliott J. Meer, Jessica M. Terry, Daniel Riordan, Niranjan Srinivas, Tarjei S. Mikkelsen, Jonathan S. Weissman, Britt Adamson
bioRxiv 503367; doi: https://doi.org/10.1101/503367
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Direct capture of CRISPR guides enables scalable, multiplexed, and multi-omic Perturb-seq
Joseph M. Replogle, Albert Xu, Thomas M. Norman, Elliott J. Meer, Jessica M. Terry, Daniel Riordan, Niranjan Srinivas, Tarjei S. Mikkelsen, Jonathan S. Weissman, Britt Adamson
bioRxiv 503367; doi: https://doi.org/10.1101/503367

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