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 April 4, 2025)
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Methods to use flow cytometry and sanger sequencing to evaluate CRISPR/Cas9 CD19 knockout in GM24385 cells
Published
Author(s)
Sarah Inwood, , , ,
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
Issues
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Created June 15, 2022, Updated March 19, 2025