Aarhus Universitets segl

English

Du er her: AU » Om AU » Organisation » CRISPR-Cas9-directed gene tagging using a single integras...

Om AU

CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

Standard

CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch. / Thomsen, Emil Aagaard; Skipper, Kristian Alsbjerg; Andersen, Sofie et al.
I: Molecular Therapy - Nucleic Acids, Bind 29, 09.2022, s. 563-576.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

Harvard

Thomsen, EA, Skipper, KA, Andersen, S, Haslund, D, Skov, TW & Mikkelsen, JG 2022, 'CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch', Molecular Therapy - Nucleic Acids, bind 29, s. 563-576. https://doi.org/10.1016/j.omtn.2022.08.005

APA

Thomsen, E. A., Skipper, K. A., Andersen, S., Haslund, D., Skov, T. W., & Mikkelsen, J. G. (2022). CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch. Molecular Therapy - Nucleic Acids, 29, 563-576. https://doi.org/10.1016/j.omtn.2022.08.005

CBE

Thomsen EA, Skipper KA, Andersen S, Haslund D, Skov TW, Mikkelsen JG. 2022. CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch. Molecular Therapy - Nucleic Acids. 29:563-576. https://doi.org/10.1016/j.omtn.2022.08.005

MLA

Thomsen, Emil Aagaard et al. "CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch". Molecular Therapy - Nucleic Acids. 2022, 29. 563-576. https://doi.org/10.1016/j.omtn.2022.08.005

Vancouver

Thomsen EA, Skipper KA, Andersen S, Haslund D, Skov TW, Mikkelsen JG. CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch. Molecular Therapy - Nucleic Acids. 2022 sep.;29:563-576. doi: 10.1016/j.omtn.2022.08.005

Author

Thomsen, Emil Aagaard ; Skipper, Kristian Alsbjerg ; Andersen, Sofie et al. / CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch. I: Molecular Therapy - Nucleic Acids. 2022 ; Bind 29. s. 563-576.

Bibtex

@article{1ecf8d39de434f268c24e27f005cc80c,
title = "CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch",
abstract = "Locus-directed DNA cleavage induced by the CRISPR-Cas9 system triggers DNA repair mechanisms allowing gene repair or targeted insertion of foreign DNA. For gene insertion to be successful, availability of a homologous donor template needs to be timed with cleavage of the DNA by the Cas9 endonuclease guided by a target-specific single guide RNA (sgRNA). We present a novel approach for targeted gene insertion based on a single integrase-defective lentiviral vector (IDLV) carrying a Cas9 off switch. Gene insertion using this approach benefits from transposon-based stable Cas9 expression, which is switched off by excision-only transposase protein co-delivered in IDLV particles carrying a combined sgRNA/donor vector. This one-vector approach supports potent (up to >80%) knockin of a full-length EGFP gene sequence. This traceless cell engineering method benefits from high stable levels of Cas9, timed intracellular availability of the molecular tools, and a built-in feature to turn off Cas9 expression after DNA cleavage. The simple technique is based on transduction with a single IDLV, which holds the capacity to transfer larger donor templates, allowing robust gene knockin or tagging of genes in a single step.",
keywords = "AAV, CRISPR-Cas9, DNA transposon, Donor template, Gene tagging, HDR, IDLV, lentivirus, MT: Delivery Strategies, piggyBac, protein delivery",
author = "Thomsen, {Emil Aagaard} and Skipper, {Kristian Alsbjerg} and Sofie Andersen and Didde Haslund and Skov, {Thomas Wisbech} and Mikkelsen, {Jacob Giehm}",
note = "Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = sep,
doi = "10.1016/j.omtn.2022.08.005",
language = "English",
volume = "29",
pages = "563--576",
journal = "Molecular Therapy - Nucleic Acids",
issn = "2162-2531",
publisher = "Nature Publishing Group",

}

RIS

TY - JOUR

T1 - CRISPR-Cas9-directed gene tagging using a single integrase-defective lentiviral vector carrying a transposase-based Cas9 off switch

AU - Thomsen, Emil Aagaard

AU - Skipper, Kristian Alsbjerg

AU - Andersen, Sofie

AU - Haslund, Didde

AU - Skov, Thomas Wisbech

AU - Mikkelsen, Jacob Giehm

N1 - Publisher Copyright: © 2022 The Author(s)

PY - 2022/9

Y1 - 2022/9

N2 - Locus-directed DNA cleavage induced by the CRISPR-Cas9 system triggers DNA repair mechanisms allowing gene repair or targeted insertion of foreign DNA. For gene insertion to be successful, availability of a homologous donor template needs to be timed with cleavage of the DNA by the Cas9 endonuclease guided by a target-specific single guide RNA (sgRNA). We present a novel approach for targeted gene insertion based on a single integrase-defective lentiviral vector (IDLV) carrying a Cas9 off switch. Gene insertion using this approach benefits from transposon-based stable Cas9 expression, which is switched off by excision-only transposase protein co-delivered in IDLV particles carrying a combined sgRNA/donor vector. This one-vector approach supports potent (up to >80%) knockin of a full-length EGFP gene sequence. This traceless cell engineering method benefits from high stable levels of Cas9, timed intracellular availability of the molecular tools, and a built-in feature to turn off Cas9 expression after DNA cleavage. The simple technique is based on transduction with a single IDLV, which holds the capacity to transfer larger donor templates, allowing robust gene knockin or tagging of genes in a single step.

AB - Locus-directed DNA cleavage induced by the CRISPR-Cas9 system triggers DNA repair mechanisms allowing gene repair or targeted insertion of foreign DNA. For gene insertion to be successful, availability of a homologous donor template needs to be timed with cleavage of the DNA by the Cas9 endonuclease guided by a target-specific single guide RNA (sgRNA). We present a novel approach for targeted gene insertion based on a single integrase-defective lentiviral vector (IDLV) carrying a Cas9 off switch. Gene insertion using this approach benefits from transposon-based stable Cas9 expression, which is switched off by excision-only transposase protein co-delivered in IDLV particles carrying a combined sgRNA/donor vector. This one-vector approach supports potent (up to >80%) knockin of a full-length EGFP gene sequence. This traceless cell engineering method benefits from high stable levels of Cas9, timed intracellular availability of the molecular tools, and a built-in feature to turn off Cas9 expression after DNA cleavage. The simple technique is based on transduction with a single IDLV, which holds the capacity to transfer larger donor templates, allowing robust gene knockin or tagging of genes in a single step.

KW - AAV

KW - CRISPR-Cas9

KW - DNA transposon

KW - Donor template

KW - Gene tagging

KW - HDR

KW - IDLV

KW - lentivirus

KW - MT: Delivery Strategies

KW - piggyBac

KW - protein delivery

U2 - 10.1016/j.omtn.2022.08.005

DO - 10.1016/j.omtn.2022.08.005

M3 - Journal article

C2 - 36090759

AN - SCOPUS:85136182555

VL - 29

SP - 563

EP - 576

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

SN - 2162-2531

ER -