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

Research output: Contribution to journalArticlepeer-review

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 languageEnglish
Pages (from-to)123-134
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Volume2770
DOIs
Publication statusPublished - 2024

Bibliographical note

© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Mice
  • Animals
  • Gene Editing/methods
  • CRISPR-Cas Systems/genetics
  • Ribonucleoproteins/metabolism
  • RNA, Guide, CRISPR-Cas Systems
  • Germ Cells/metabolism

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