Lipofection-Based Delivery of CRISPR/Cas9 Ribonucleoprotein for Gene Editing in Male Germline Stem Cells

Mariella Obermeier; Vera Rogiers; Tamara Vanhaecke; Yoni Baert

doi:10.1007/978-1-0716-3698-5_10

Lipofection-Based Delivery of CRISPR/Cas9 Ribonucleoprotein for Gene Editing in Male Germline Stem Cells

Mariella Obermeier, Vera Rogiers, Tamara Vanhaecke, Yoni Baert

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Abstract

Gene editing in the murine germline is a valuable approach to investigate germ cell maturation and generate mouse models. Several studies demonstrated that CRISPR/Cas9 alters the genome of cultured male mouse germline stem cells delivered by electroporation of plasmids. Recently, we showed proof-of-principle that gene knockout can be effectively targeted in mouse germline stem cells by lipofecting Cas9:gRNA ribonucleoproteins. In this protocol, we describe a simple, fast, and cheap workflow for gene editing via the lipofection of non-integrative ribonucleoproteins in murine male germline stem cells.

Original language	English
Pages (from-to)	123-134
Number of pages	12
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	2770
DOIs	https://doi.org/10.1007/978-1-0716-3698-5_10
Publication status	Published - 2024

Bibliographical note

Keywords

CRISPR/Cas9
Germline stem cell
Gene editing
Ribonucleoprotein
Germline genome editing
Gene knockout
Spermatogonial stem cell
Oligodeoxynucleotide

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10.1007/978-1-0716-3698-5_10

Obermeier et al 2024_Yoni Baert_MB_03082023_2Accepted author manuscript, 309 KBLicence: Unspecified

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title = "Lipofection-Based Delivery of CRISPR/Cas9 Ribonucleoprotein for Gene Editing in Male Germline Stem Cells",

abstract = "Gene editing in the murine germline is a valuable approach to investigate germ cell maturation and generate mouse models. Several studies demonstrated that CRISPR/Cas9 alters the genome of cultured male mouse germline stem cells delivered by electroporation of plasmids. Recently, we showed proof-of-principle that gene knockout can be effectively targeted in mouse germline stem cells by lipofecting Cas9:gRNA ribonucleoproteins. In this protocol, we describe a simple, fast, and cheap workflow for gene editing via the lipofection of non-integrative ribonucleoproteins in murine male germline stem cells.",

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doi = "10.1007/978-1-0716-3698-5_10",

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T1 - Lipofection-Based Delivery of CRISPR/Cas9 Ribonucleoprotein for Gene Editing in Male Germline Stem Cells

AU - Obermeier, Mariella

AU - Rogiers, Vera

AU - Vanhaecke, Tamara

AU - Baert, Yoni

PY - 2024

Y1 - 2024

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AB - Gene editing in the murine germline is a valuable approach to investigate germ cell maturation and generate mouse models. Several studies demonstrated that CRISPR/Cas9 alters the genome of cultured male mouse germline stem cells delivered by electroporation of plasmids. Recently, we showed proof-of-principle that gene knockout can be effectively targeted in mouse germline stem cells by lipofecting Cas9:gRNA ribonucleoproteins. In this protocol, we describe a simple, fast, and cheap workflow for gene editing via the lipofection of non-integrative ribonucleoproteins in murine male germline stem cells.

KW - CRISPR/Cas9

KW - Germline stem cell

KW - Gene editing

KW - Ribonucleoprotein

KW - Germline genome editing

KW - Gene knockout

KW - Spermatogonial stem cell

KW - Oligodeoxynucleotide

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JF - Methods in molecular biology (Clifton, N.J.)

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Lipofection-Based Delivery of CRISPR/Cas9 Ribonucleoprotein for Gene Editing in Male Germline Stem Cells

Abstract

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FWOAL1081: Organoids from transgender women testes: from medical waste to a promising testicular toxicity assay.

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Lipofection-Based Delivery of CRISPR/Cas9 Ribonucleoprotein for Gene Editing in Male Germline Stem Cells

Abstract

Bibliographical note

Keywords

Access to Document

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Other files and links

Embargoed Document

Fingerprint

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FWOAL1081: Organoids from transgender women testes: from medical waste to a promising testicular toxicity assay.

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