Methods to use flow cytometry and sanger sequencing to evaluate CRISPR/Cas9 CD19 knockout in GM24385 cells

Published

June 15, 2022

Author(s)

Sarah Inwood, Linhua Tian, Kirsten Parratt, Samantha Maragh, Lili Wang

Abstract

CRISPR/Cas9, one of the most commonly used genome editing systems, has been used for production of protein, cell and gene therapies. While an exciting tool, there is not a lot of long-term data about its genomic and phenotypic stability and off-target effects that may arise during the editing process. Since the whole cell is used in the patient with cell and gene therapies, cell characterization is essential for product safety. In support of the production of safe and effective CRISPR/Cas9 engineered protein, cell and gene therapies, the objective of this work is to describe a protocol for measuring the edits from CRISPR/Cas9 using flow cytometry on a B-lymphoblast cell line, GM24385, whose genome sequence has been well characterized.

Citation

Biotechniques

Pub Type

Journals

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Keywords

gene editing, genome editing, CRISPR/Cas9, flow cytometry, GM24385, CD19

Citation

Inwood, S. , Tian, L. , Parratt, K. , Maragh, S. and Wang, L. (2022), Methods to use flow cytometry and sanger sequencing to evaluate CRISPR/Cas9 CD19 knockout in GM24385 cells, Biotechniques, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=933798 (Accessed October 17, 2025)