CRISPR/Cas9 Genome Editing in Embryonic Stem Cells.
Targeted mutagenesis is required to evaluate the function of DNA segments across the genome. In recent years the CRISPR/Cas9 technology has been widely used for functional genome studies and is partially replacing classical homologous recombination methods in different aspects. CRISPR/Cas9-derived tools indeed allow the production of a wide-range of engineered mutations: from point mutations to large chromosomal rearrangements such as deletions, duplications and inversions. Here we present a protocol to engineer Embryonic Stem Cells (ESC) with desired mutations using transfection of custom-made CRISPR/Cas9 vectors. These methods allow the in vivo modeling of congenital mutations and the functional interrogation of DNA sequences.
Development and Disease Group, Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, Berlin, 14195, Germany.
Guillaume
G
Andrey
Development and Disease Group, Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, Berlin, 14195, Germany. spielman@molgen.mpg.de.
Malte
M
Spielmann
D000818:Animals; D064113:CRISPR-Cas Systems; D000072669:Gene Editing; D005818:Genetic Engineering; D051379:Mice; D008957:Models, Genetic; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology*; D009154:Mutation; D014162:Transfection
CRISPR; Cas9; ES cells; Genome engineering; Indels; Mouse; Point mutation; Structural variants