The mission of the UTSW Transgenic Core is to create, propagate, and preserve lines of genetically engineered rodents. The Core's staff generate conventional transgenic mice and rats from various inbred backgrounds using either single or multiple small transgenes and BACs. The Core has been creating genome edited rodents for ~ 10 years using the latest CRISPR/Cas9 editing tools. These efforts include creating simple KO’s, KI’s using oligo, megamer, HDR block, or plasmid-based templates for homologous recombination (HR), and the creation of conditional alleles using either 2 oligo templates containing LoxP motifs or a single long ssDNA template with the appropriate LoxP motifs. The Core's staff continue to perform gene targeting in mouse embryonic stem (ES) cells on a small scale, expansion of in-house targeted ES cell clones and consortium-derived clones, and the preparation of select targeted ES cell clones for injection into blastocysts to create chimeric mice.
The Core staff routinely provide UTSW Investigators access to a wide array of assisted reproductive techniques. These include the cryopreservation of sperm, 2-cell embryos and ovaries from select lines of mice. The frozen gametes are stored in the Core’s cryopreservation facility on the South Campus and can be distributed to Investigators world-wide. Core staff can reconstitute lines of mice from cryopreserved gametes by either thawing embryos and transferring them to surrogate females or by thawing sperm, performing in vitro fertilization followed by embryo transfers. The Core staff routinely rederive lines of mice from conventional housing to an SPF barrier status. If the need arises, the Core can rescue a line of mice that has reached reproductive senescence or propagate a line of mice so that a very large number of animals can be produced in a single day.
Services Offered
Protocols
Robert Hammer, Ph.D. | Director
robert.hammer@utsouthwestern.edu
214-645-6292
Hours | Location |
Monday to Friday: 8am-5pm |
6001 Forest Park Road |
Name | Role | Phone | Location | |
---|---|---|---|---|
John Ritter and Mylinh Nguyen |
214-645-6288
|
john.ritter@utsouthwestern.edu; mylinh.nguyen@utsouthwestern.edu
|
ND11.223
|
Price list |
► CRISPR (18) | |||
Name | Description | Price | |
---|---|---|---|
KnockOut - INDEL/Exon loss - C57BL/6N mouse eggs / per 200 embryos |
Delivery of CRISPR reagents into C57BL/6N mouse eggs |
Inquire | |
KnockOut - INDEL/Exon loss - C57BL/6J mouse eggs / per 200 embryos |
Delivery of CRISPR reagents into C57BL/6J mouse eggs |
Inquire | |
KnockIn - Oligo 'less than' 200bp ssODN - C57BL/6N mouse eggs / per 300 embryos |
Delivery of CRISPR reagents into C57BL/6N mouse eggs |
Inquire | |
KnockIn - Oligo 'less than' 200bp ssODN - C57BL/6J mouse eggs / per 300 embryos |
Delivery of CRISPR reagents into C57BL/6J mouse eggs |
Inquire | |
KnockIn - Megamer 'greater than' 200bp ssODN - C57BL/6N mouse eggs / per 400 embryos |
Delivery of CRISPR reagents into C57BL/6N mouse eggs |
Inquire | |
KnockIn - Megamer 'greater than' 200bp ssODN - C57BL/6J mouse eggs / per 400 embryos |
Delivery of CRISPR reagents into C57BL/6J mouse eggs |
Inquire | |
KnockIn - Conditional - ssODN x2 - C57BL/6N mouse eggs / per 400 embryos |
Delivery of CRISPR reagents into C57BL/6N mouse eggs |
Inquire | |
KnockIn - Conditional - ssODN x2 - C57BL/6J mouse eggs / per 400 embryos |
Delivery of CRISPR reagents into C57BL/6J mouse eggs |
Inquire | |
KnockIn - Plasmid dsDNA - C57BL/6N mouse eggs / per 400 embryos |
Delivery of CRISPR reagents into C57BL/6N mouse eggs |
Inquire | |
KnockIn - Plasmid dsDNA - C57BL/6J mouse eggs / per 400 embryos |
Delivery of CRISPR reagents into C57BL/6J mouse eggs |
Inquire | |
Knock-In - HDR-Block from IDT - C57BL/6N mouse eggs / per 400 embryos |
Alt-R HDR Block from IDT is a double strand DNA template, 201 to 3000bp in length, that is used to generate a CRISPR Knock-In's. |
Inquire | |
Knock-In - HDR-Block from IDT - C57BL/6J mouse eggs / per 400 embryos |
Alt-R HDR Block from IDT is a double strand DNA template, 201 to 3000bp in length, that is used to generate a CRISPR Knock-In's in the C57BL/6J mouse |
Inquire | |
IVF Generated Embryos and CRISPR Injections |
PI should request this service when the use of an established line of mice is needed to further generate gene edited rodents using CRISPR reagents.
|
Inquire | |
Rescue Conditional KO - Insert missing 5' or 3' LoxP Site |
The Core will generate embryos from the PI mouse line using IVF and then inject CRISPR reagents to insert the missing LoxP site. |
Inquire | |
CRISPR-modified Rats - KO / per 200 embryos |
Delivery of CRISPR reagents into Rat eggs |
Inquire | |
CRISPR-modified Sprague Dawley Rats - KI or KO / per 300 embryos |
Delivery of CRISPR reagents into Sprague Dawley rat eggs. |
Inquire | |
CRISPR-modified Brown Norway Rats - KI or KO / per 300 embryos |
Delivery of CRISPR reagents to rat embryos from the Brown Norway strain. |
Inquire | |
Other Strain Fee |
This fee will cover the cost of ordering a strain not routinely in use by TTC and any associated reagents, supplies, or per diem. |
Inquire | |
► Transgenic Rodents (6) | |||
Name | Description | Price | |
Microinjection into the mouse pronucleus - C57Bl/6NCrL (Charles River #027) |
Microinjection of purified DNA and embryo transfers of 300 pronucleus stage C57BL/6N embryos |
Inquire | |
Microinjection into the mouse pronucleus - C57Bl/6J (Jackson Labs #000664) |
Microinjection of purified DNA and embryo transfers of 300 pronucleus stage C57BL/6J embryos |
Inquire | |
Microinjection into the mouse pronucleus - B6SJLF1 (Jackson Labs #100012) |
Microinjection of purified DNA and embryo transfers of 300 pronucleus stage B6SJLF1 embryos |
Inquire | |
Transient Transgenic / Developmental Assay Service |
One day Injections of DNA Shipment of Recipent Females for eDay Assay C57Bl/6N strain
|
Inquire | |
Isolation and Purification of the Transgene Fragment from the Parent Plasmid |
Isolation of the transgene fragment from the parent plasmid and purification for microinjection |
Inquire | |
Propagation of Plasmid or BAC and Purification of cirDNA |
Plasmid or BAC Propagation and DNA purification for microinjections |
Inquire | |
► Assisted Reproductive Techniques (ART) (6) | |||
Name | Description | Price | |
Sperm recovery, in vitro fertilization, and embryo transfer / per 2 – 4 transfers |
The Core will thaw the cryopreserved sperm, perform in vitro fertilization, culture the eggs to the 2-cell stage, and transfer the embryos to pseudopregnant recipients |
Inquire | |
Embryo recovery and transfer / per 2 – 4 transfers |
Thaw and recover cryopreserved embryos and transfer them to pseudopregnant recipients |
Inquire | |
Ovary recovery and transplantation / per 2 – 4 transfers |
Fresh or cryopreserved bisected ovaries and surgically transfer the ovary to histocompatible recipients |
Inquire | |
Embryo Collection (only available strains) / per line |
Harvest embryo from PI mated females or strains available in the Core |
Inquire | |
Timed Matings |
The Core will set up natural matings with Core studs and females, check for a copulatory plugs, hold the successful matings till the day specified by the PI. |
Inquire | |
Tissue Collection |
The PI receives disected organs from a recently euthanized embryo donor |
Inquire | |
► Cryopreservation (8) | |||
Name | Description | Price | |
Cryopreservation of Rat Embryos |
1.The Transgenic Core Center will cryopreserve fertilized eggs from transgenic and knock-out strains of rat. The Core will freeze down at least 150 embryos per line. 2. The embryos will be placed in vials of 50 embryos/vial. 3. The PI will have an annual storage fee, starting the following year, to cover the cost of LN2 supplied to Core dewars. 4. Optional freeze and shipment can be requested. 5. The investigator is responsible for initiating and coordinating all transfer of animal requests to the Core. 6. The investigator will provide the Center with 7 – 8 proven stud males 36 weeks of age, the males' birthdate, genetic background, and genotype. Following the cryopreservation, the males can be returned to the PI's animal rooms, if they are at a LEVEL 2 health status or lower. |
Inquire | |
Cryopreservation – Mouse Embryos |
This service will provide the PI cryopreserved for their transgenic and knock-out strains of mice at approximately 200 fertilized eggs and 35 eggs/vial. |
Inquire | |
Cryopreservation – Mouse Sperm |
This service will provide the PI with cryopreserve sperm from their transgenic and knock-out strains of mice 1. 10 µl/straw with an estimate of 8-10 straws per male |
Inquire | |
Cryopreservation of Sperm for Shipment |
The PI will provide 1 proven stud males to the Core in order to perform cryopreservation of a mouse line for the purpose of shipping to another institution. |
Inquire | |
Cryopreservation – Mouse Ovaries (bisected) |
This service will provide the PI with cryopreserved, bisected ovaries from their transgenic and knock-out strains of mice. 1. Both ovaries are bisected and stored as a half per vial. 2. Up to 3 females per line. |
Inquire | |
Cryopreservation of Rat Sperm |
This service will provide the PI cryopreserve sperm from their transgenic and knock-out strains of mice; 10 µl/straw / per 8-10 straws / male |
Inquire | |
Storage / per 1 year |
Annual Storage Fee - after the first year of cryopreservation either a line for sperm, embryo, or ovary. |
Inquire | |
Vapor Shipper - LN2 Charge |
Fee to cover the cost of adding LN2 to a dry-shipper in preparation of gamate transfer. |
Inquire | |
► Rederivation (1) | |||
Name | Description | Price | |
Rederivation of Mouse Line |
Rederive 2-3 litters from contaminated line into a SPF status |
Inquire |