Mouse Gene Manipulation

Children's Hospital Intellectual and Developmental Disabilities Research Center

Center URL: https://iddrc.org/
Core URL: http://www.iddrc.org/cores/gene-manipulation/

Core Personnel
Core Director: Clifford Woolf, MB, BCh, PhD
Core Manager: Mantu Bhaumik, PhD

Core Description

The Mouse Gene Manipulation Core centralizes the production of genetically altered mice for IDDRC investigators in a cost-effective and efficient manner. The Core also provides expert assistance in the additional areas required for generating genetically altered mice, including transgene and targeting construct production, and transgenic mouse colony management. Core personnel are available to train investigators in all of the techniques required for the generation of genetically modified mice.

Services

Genome editing in vivo using CRISPR/Cas9
Transgenic DNA microinjection
Gene targeting by homologous recombination
Blastocyst injection to produce chimeric mice
Cryopreservation of mouse strains
Rederivation by IVF and embryos transfer
Derivation of ES cell line from mutant strain
Genotyping founders by Southern blot

Equipment

Microinjection Equipment

Pronuclear and blastocyst injection setup consisting of:
o one Nikon Diaphot inverted microscope with 20X phase, 4X plan, and 10,20, and 40X DIC objectives
o one video camera with monitor hooked to the Nikon for teaching
o two Eppendorf Transferman NK micromanipulats
o two fine hydraulic Narishige micromanipulators (MO-188)
o one Narishige pressure injector, Model IM-200
o one vibration isolation table

Pronuclear and blastocyst injection setup consisting of:
o one Nikon TE2000-S inverted microscope with 20X phase, 4X plan, and 20X DIC objectives
o two Eppendorf Transferman 4r micromanipulators
o one Eppendorf femtojet micro pressure injector
o one PrimeTech Piezo drill
o one vibration isolation table

Additional microinjection and surgical equipment includes:
o two Nikon SMZ-U dissecting microscopes
o one Nikon double headed teaching dissecting microscope
o one Flaming/Brown P87 horizontal pipet puller
o one Kopf 720 vertical pipet puller
o one Defonbrune-type microforge with stereo zoom optics
o one Narishige EG-4 needle grinder
o one C02 incubator RMI3000S-7-AB4
o one slide warmer for waking anesthetized animals
o one BTX ECM 2001 electro cell manipulator system

ES Cell Culture Equipment
o two C02 incubators for growth of ES cells
o two Baker Biological Safety cabinets
o one auto fill liquid nitrogen storage container
o one Nikon TMS inverted tissue culture microscope
o one Biorad electroporation gene pulser with capacitance extender
o One Sanyo -80 freezer

Molecular Biology Equipment
o one MJ Research PCR machine
o one Eppendorf microcentrifuge
o one Eppendorf centrifuge 5702R
o one ThermoScientific -80/TSU series
o one water bath
o one heat block
o one shaker
o one vortexer
o two 20x25 cm horizontal electrophoresis systems
o two 13x16 cm horizontal electrophoresis system
o two 7x10 cm horizontal electrophoresis system
o one Techne hybridization oven

Cryopreservation
o one BioCool-IV controlled rate freezer
o one micro-Osmette osmometer
o one Thermo Scientific CryoPlus2 liquid nitrogen freezer
o one Leica M205C microscope & MC170HD camera
o one IMV SYMS-III straw sealer

Computer Equipment
o four iMac
o one hp LaserJet M252dw