Mouse Gene Manipulation Core
The Mouse Gene Manipulation & Genome Editing Core Facility provides services to the investigators to create genetically engineered mouse models. Our goal is to offer expert advice and technical expertise to the investigators towards the various approaches in achieving desired mouse model in an easily accessible, affordable and efficient manner.
The core facility consists of a director, an assistant director and trained staff members. The assistant director oversees daily operation including expert consultation, quality control of regents, monitor progress of each project, communicating with the PI and lab members, trouble shooting, scheduling, billing and direct supervision of staff members.
We alter mouse genome either by utilizing genome editing CRISPR/Cas9 reagents, random insertion of transgenic DNA or gene targeting techniques in mouse embryonic stem cells. Core also provides services towards the identification, propagation and management of founder mouse lines, cryopreservation, re-derivation and development of mouse-oriented research projects in the IDDRC and Boston Children’s Hospital scientific community.
Mouse Genome editing technique relies on recently developed genome editing tools using CRISPR /Cas9 system. These reagents are used to introduce precise modification of mouse genome, allowing efficient founder generation in a single step. The genome editing method can be used both in vitro and in vivo to create autosomal or germ line modifications. CRISPR/Cas9 reagents allow specific modification from a single nucleotide change to insertion or deletion of large DNA fragments at a specific target site of mouse genome. The relative ease and accessibility of CRISPR/Cas9 reagents, lower cost and higher efficiency is an attractive approach in generating desired mouse model. Learn about mouse genome editing services
Gene targeting - Gene specific mutation is created by introducing DNA constructs harboring desired modification of the gene or genes into pluripotent mouse embryonic stem (mES) cells via homologous recombination. This technique relies on homology dependent DNA recombination thus require homology arms spanning gene of interest. The gene targeted mES cells are then introduced into blastocysts to integrate pluripotent mES cells harboring mutations into inner cell mass (ICM) to develop into a chimeric or mosaic mouse. Creation of chimeric founder mouse bearing specific modification is capable of transmitting mutation through germ cells to the next generation as heritable mutation. Users can choose mES cell lines from two congenic (C57 and 129Sv) and a hybrid (C57x129Sv) genetic background for targeting experiments or any imported mES cell lines. Learn about gene targeting services
Transgenic mouse models are generated by using DNA constructs with desired modification/s that is introduced into fertilized one-cell embryos to achieve random integration into mouse genome. This method typically is a gain-of-function to study gene of interest in vivo, which is expressed under the control of a tissue specific or ubiquitous promoter allowing study of a gene function in space and time. We offer founder generation using C57Bl/6 as standard strain; however, users can choose a specific strain of mice or mutant strain for additional fees. Learn about transgenic mouse services