Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology

Harald Lahm; Niraj K. Singh; Irina Gottmann; Stefanie A. Doppler; Elda Dzilic; Heike Preisler; Stephanie Schneider; Rüdiger Lange; Markus Krane; Martina Dreßen

doi:10.1016/j.scr.2023.103261

Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology

Harald Lahm, Niraj K. Singh, Irina Gottmann, Stefanie A. Doppler, Elda Dzilic, Heike Preisler, Stephanie Schneider, Rüdiger Lange, Markus Krane, Martina Dreßen

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Abstract

The identification of TBX5-related regulatory sequences in genes essential for heart development is hampered by the absence of antibodies which allow precipitation of TBX5:DNA complexes. Employing CRISPR/Cas9 technology, we have inserted a FLAG-tag sequence at the end of exon 9 of the TBX5 gene prior to the stop codon by homologous recombination. The translated TBX5-FLAG fusion protein of the three iPSC lines can effectively be precipitated by anti-FLAG antibodies and, thus, allow the detection of specific TBX5-binding sites and their associated genes.

Original language	English
Article number	103261
Journal	Stem Cell Research
Volume	74
DOIs	https://doi.org/10.1016/j.scr.2023.103261
State	Published - Feb 2024

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Lahm, H., Singh, N. K., Gottmann, I., Doppler, S. A., Dzilic, E., Preisler, H., Schneider, S., Lange, R., Krane, M., & Dreßen, M. (2024). Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology. Stem Cell Research, 74, Article 103261. https://doi.org/10.1016/j.scr.2023.103261

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title = "Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology",

abstract = "The identification of TBX5-related regulatory sequences in genes essential for heart development is hampered by the absence of antibodies which allow precipitation of TBX5:DNA complexes. Employing CRISPR/Cas9 technology, we have inserted a FLAG-tag sequence at the end of exon 9 of the TBX5 gene prior to the stop codon by homologous recombination. The translated TBX5-FLAG fusion protein of the three iPSC lines can effectively be precipitated by anti-FLAG antibodies and, thus, allow the detection of specific TBX5-binding sites and their associated genes.",

author = "Harald Lahm and Singh, {Niraj K.} and Irina Gottmann and Doppler, {Stefanie A.} and Elda Dzilic and Heike Preisler and Stephanie Schneider and R{\"u}diger Lange and Markus Krane and Martina Dre{\ss}en",

note = "Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2024",

month = feb,

doi = "10.1016/j.scr.2023.103261",

language = "English",

volume = "74",

journal = "Stem Cell Research",

issn = "1873-5061",

publisher = "Elsevier B.V.",

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Lahm, H, Singh, NK, Gottmann, I, Doppler, SA, Dzilic, E, Preisler, H, Schneider, S, Lange, R , Krane, M & Dreßen, M 2024, 'Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology', Stem Cell Research, vol. 74, 103261. https://doi.org/10.1016/j.scr.2023.103261

TY - JOUR

T1 - Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology

AU - Lahm, Harald

AU - Singh, Niraj K.

AU - Gottmann, Irina

AU - Doppler, Stefanie A.

AU - Dzilic, Elda

AU - Preisler, Heike

AU - Schneider, Stephanie

AU - Lange, Rüdiger

AU - Krane, Markus

AU - Dreßen, Martina

PY - 2024/2

Y1 - 2024/2

N2 - The identification of TBX5-related regulatory sequences in genes essential for heart development is hampered by the absence of antibodies which allow precipitation of TBX5:DNA complexes. Employing CRISPR/Cas9 technology, we have inserted a FLAG-tag sequence at the end of exon 9 of the TBX5 gene prior to the stop codon by homologous recombination. The translated TBX5-FLAG fusion protein of the three iPSC lines can effectively be precipitated by anti-FLAG antibodies and, thus, allow the detection of specific TBX5-binding sites and their associated genes.

AB - The identification of TBX5-related regulatory sequences in genes essential for heart development is hampered by the absence of antibodies which allow precipitation of TBX5:DNA complexes. Employing CRISPR/Cas9 technology, we have inserted a FLAG-tag sequence at the end of exon 9 of the TBX5 gene prior to the stop codon by homologous recombination. The translated TBX5-FLAG fusion protein of the three iPSC lines can effectively be precipitated by anti-FLAG antibodies and, thus, allow the detection of specific TBX5-binding sites and their associated genes.

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U2 - 10.1016/j.scr.2023.103261

DO - 10.1016/j.scr.2023.103261

M3 - Article

C2 - 38100916

AN - SCOPUS:85179820992

SN - 1873-5061

VL - 74

JO - Stem Cell Research

JF - Stem Cell Research

M1 - 103261

ER -

Insertion of a FLAG-tag sequence at the end of exon 9 of the TBX5 gene in three induced pluripotent stem cell lines (DHMi004-A-4, DHMi004-A-5, DHMi004-A-6) by CRISPR/Cas9 technology

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