Precise and Scarless Insertion of Transposable Elements by Cas9-Mediated Genome Engineering - Archive ouverte HAL
Chapitre D'ouvrage Année : 2023

Precise and Scarless Insertion of Transposable Elements by Cas9-Mediated Genome Engineering

Résumé

Transposable element insertions can have broad effects on gene expression, ranging from new regulatory functions to pathogenic consequences by transplanting new cis-regulating elements or perturbing existing ones. Genetic manipulation of such DNA sequences can help decipher their mechanism of action. Here, we describe a CRISPR-Cas9-mediated two-step approach to precisely insert transposable elements into into the genome of cultured human cells, without scar or reporter gene. First, a double-selection cassette is inserted into the desired target locus. Once a clone containing a single copy of this cassette has been isolated, a second editing step is performed to exchange the double-selection cassette with a markerless transposable element sequence. More generally, this method can be used for knocking in any large insert without genetic markers.
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Dates et versions

hal-04246145 , version 1 (17-10-2023)

Identifiants

Citer

Vivien Marie Weber, Aurélien J. Doucet, Gael Cristofari. Precise and Scarless Insertion of Transposable Elements by Cas9-Mediated Genome Engineering. Miguel R. Branco; Alexandre de Mendoza Soler. Transposable Elements, 2607, Springer US, pp.329-353, 2023, Methods in Molecular Biology, ⟨10.1007/978-1-0716-2883-6_15⟩. ⟨hal-04246145⟩
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