Transgenic Mouse Services, transgene knockin

Introduction

GeneCopeia provides transgenic mouse services using CRISPR-Cas9. Many applications are readily within reach, including constitutive and conditional gene knockout, mutagenesis, correction of disease mutations, transgene knockin, and in-frame fusion tagging.

Advantages

Track record of success (Figure 1).
Fast mouse line creation using CRISPR-Cas9 and pronuclear injection (Figure 2). F1 modified mice in as little as 6 months.
In-depth project consultation.
Expert CRISPR clone design, construction, and mouse injection
Compatible with GeneCopoeia's GeneHero™ ROSA26 Safe Harbor knockin clones.

Case study: Gene tagging using GeneCopoeia's CRISPR transgenic mouse services

Figure 1. In-frame GFP fusion of a chromosomal gene in mice using GeneCopoeia CRISPR technology. Cas9 mRNA, sgRNA, and a donor plasmid targeting the M. musculus P2xr2 gene (NCBI gene ID: 231602) were injected into 1-cell stage mouse embryos. CRISPR-Cas9 stimulates homologous recombination between the unmodified chromosome and the homologous donor, resulting in an in-frame, 3’ terminal GFP-tagged gene. Positive founder mice were identified by PCR and sequencing. Gray boxes: P2xr2 exons.

Figure 2. Site-specific genome editing mediated by TALEN or CRISPR-Cas9, as well as its application on transgenic mouse generation.

References

1. Knockout mice by TALEN-mediated gene targeting.Nature Biotrchnology.2012,31,23-24

2. Generation of gene-modified mice via Cas9/RNA-mediated gene targeting.Cell Research.2013,23,720-723.

3. One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Geneme Engineeing.Cell 2013,153,1-9.