FACS-Based Sequencing Approach to Evaluate Cell Type to Genotype Associations Using Cerebral Organoids

Liam Murray; Meagan N Olson; Nathaniel Barton; Pepper Dawes; Yingleong Chan; Elaine T Lim

doi:10.1007/978-1-0716-3287-1_15

FACS-Based Sequencing Approach to Evaluate Cell Type to Genotype Associations Using Cerebral Organoids

Methods Mol Biol. 2023:2683:193-199. doi: 10.1007/978-1-0716-3287-1_15.

Authors

Liam Murray^{1

2

3

4}, Meagan N Olson^{1

2

3

4}, Nathaniel Barton^{1

2

3

4}, Pepper Dawes^{1

2

3

4}, Yingleong Chan^{1

2

3}, Elaine T Lim^{5

6

7

8}

Affiliations

¹ Program in Bioinformatics and Integrative Biology, University of Massachusetts Chan Medical School, Worcester, MA, USA.
² Department of Neurology, University of Massachusetts Chan Medical School, Worcester, MA, USA.
³ NeuroNexus Institute, University of Massachusetts Chan Medical School, Worcester, MA, USA.
⁴ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA, USA.
⁵ Program in Bioinformatics and Integrative Biology, University of Massachusetts Chan Medical School, Worcester, MA, USA. Elaine.Lim@umassmed.edu.
⁶ Department of Neurology, University of Massachusetts Chan Medical School, Worcester, MA, USA. Elaine.Lim@umassmed.edu.
⁷ NeuroNexus Institute, University of Massachusetts Chan Medical School, Worcester, MA, USA. Elaine.Lim@umassmed.edu.
⁸ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA, USA. Elaine.Lim@umassmed.edu.

PMID: 37300776
DOI: 10.1007/978-1-0716-3287-1_15

Abstract

Recent technological developments have led to widespread applications of large-scale transcriptomics-based sequencing methods to identify genotype-to-cell type associations. Here we describe a fluorescence-activated cell sorting (FACS)-based sequencing method to utilize CRISPR/Cas9 edited mosaic cerebral organoids to identify or validate genotype-to-cell type associations. Our approach is high-throughput and quantitative and uses internal controls to enable comparisons of the results across different antibody markers and experiments.

Keywords: Cerebral organoids; FACS; Genotype to cell type; Sequencing; oFlowSeq.

FACS-Based Sequencing Approach to Evaluate Cell Type to Genotype Associations Using Cerebral Organoids

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