¹û½´ÊÓƵ

To deliver gene targeting capabilities including the production of transgenic mouse models.

Our primary purpose is generation of new mouse models for all research groups at Babraham. We specialize in zygotic gene editing which uses lower resources and has a faster turnaround (6 months for validated F1 mice). With zygotic targeting we can create a variety of mouse models such as traditional KO, deletions, point mutations, floxed and Cre-based mouse models. We can also produce more advanced models such genes tagged with Halotag, MiniTurbo, FKBP12F36V and NanoBiT. Further, we can also swap genomic DNA with other organisms to create chimeric DNA mouse models. We can also insert larger DNA sequences (>10kb) into safe harbour sites such as Rosa26 using landing pad technology. Thus, we provide a comprehensive range of mouse models for the institute.

We also provide expertise in:

• 1:1 guidance on using CRISPR/Cas9 to genome engineer your own cell line.
• Designing genotyping strategies and troubleshooting pipelines for cell lines.
• Next generation sequencing (NGS) of CRISPR/Cas9 indels and homology directed repair (HDR)
• Transient Blastocyst KO generation and developmental assays with genotyping. 
• Use of attb/attp landings site to insert large DNA fragments into the genome.
• Mouse embryonic stem cell injection into early embryos to create chimeric mice.
• Cytoplasmic injection to inject Trim-away into zygotes for protein deletion.
• Immunosurgery to isolate cells from the inner cell mast.
• Removal of loxp/Frt sites using Cre/Flip mRNA via electroporation.
• Creating multiplex mosaic mouse embryos with various libraries.
• 2-cell embryo manipulation/editing to force mosciasm.
• Use of fusion Cas9 proteins to enhance the likelihood of HDR
• QX200 for digital droplet PCR (ddPCR) applications for copy number determination, gene expression determination without CT values and clone screening applications

Contact:

Asif Nakhuda - Gene Targeting Specialist

asif.nakhuda@babraham.ac.uk