Queensland Facility for Advanced Genome Editing

QFAGE provides expert genetic modification services to researchers in the life sciences and biomedicine.

Advances in CRISPR/Cas9 genome editing technology are providing new and rapid means to develop genetic models for research. In charge of the facility are Professor Peter Koopman, overseeing mouse genome editing, and Dr Nathan Palpant, overseeing cell line genome editing. The facility has two full-time managers with extensive experience and dedication to experimental success.  

Genome editing services

Standard CRISPR/Cas9 and TG mouse production service involves three staged modules:

  • Module 1: pre-experiment design and reagents
  • Module 2: microinjection, transfer and agistment to weaning stage
  • Module 3: post-weaning analysis to handover stage

Standard CRISPR/Cas9 and cell line production service involves three staged modules:

  • Module 1: pre-experiment design, reagent preparation, and cell line QC
  • Module 2: transfection and pool efficiency analysis
  • Module 3: Single colony generation, analysis and handover stage

In general, we provide the following types of services (the cost for the standard service varies according to the type of experiment involved – please contact us for detailed pricing).

Type 1 CRISPR/Cas9 project (simple): Gene knockout or large deletion. This normally involves targeting two sgRNAs either side of the region or exon to be deleted, and no repair template, so the deletion results from non-homologous end joining (NHEJ).

Type 2 CRISPR/Cas9 project (intermediate): Generation of small, specific changes (e.g. point mutations, small specific insertions/deletions up to 60 bp, or epitope tagging). This normally involves using two sgRNAs targeted at the site of interest, and homology-director repair (HDR) via a single-stranded DNA oligonucleotide substrate (~150 bases).

Type 3 CRISPR/Cas9 project (complex): Generation of conditional (“floxed”) alleles or insertion of large DNA fragments. These experiments are much more complicated and involve either several sgRNAs (two for each LoxP insertion site) and repair templates, or a large plasmid DNA substrate (5–10 kb) to either “flox” an exon or “knock-in” a reporter or other large piece of sequence.

Other services: A standard mouse TG project involves a simplified service module 1 (preparation of injectables), plus full service modules 2 and 3. A package consisting of a customised combination of service modules for mouse or cell line production is available on request.