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Protocol for Generating Monoclonal Cell Lines with Specific Chromosomal Rearrangements

Year: 2026

Type: preprint

Abstract: Chromosomal translocations are a hallmark of hematological malignancies. This protocol enables the expeditious production of monoclonal cell lines with specific rearrangements to serve as leukemia cell models. The approach employs Cas9 to induce chromosomal breaks leading to rearrangements, followed by hierarchical subcloning and PCR-based screening to isolate and expand single-cell clones. Using the lymphoblastoid cell line RPMI8866 and the KMT2A::AFDN translocation as an example, we detail steps from sgRNA design to validation of clones with the desired translocation.

Source: Preprints.org

DOI: 10.20944/preprints202610.2438.v1

Authors: Nikolai A. Lomov, Viktoriia O. Ivanova, Vladimir S. Viushkov, Nikolai A. Nikolaev, Ekaterina V. Morgatskaia, Elena A. Zerkalenkova, Yegor S. Vassetzky, Mikhail A. Rubtsov

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: green

OpenAlex ID: W7223961939

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