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Services

Assisted Reproductive Technologies

Embryos cryopreservation

Embryos will be frozen at the two-cell stage. As part of our quality control (QC), the GEMF will thaw one straw of frozen embryos and culture them in vitro to the blastocyst stage to ensure future reviving of the embryos.

In Vitro Fertilization (IVF)

In Vitro Fertilization (IVF) will be performed from fresh, cold or frozen sperm to either generate live pups or to freeze embryos. A minimum of two recipient females will be implanted for reviving; otherwise the embryos will be frozen for archiving.

Embryos implants

Frozen embryos will be implanted into recipient females to revive a Genetically Modified (GM) mouse line. A minimum of two recipient females will be implanted.

Blastocysts injection

Injection of embryonic stem cells (ESC) or induced pluripotent stem cells (iPSC) into blastocysts to generate chimeric mice or study the contribution of cells to the developing embryos. A minimum of two recipient females will be implanted to generate chimeras.

Rederivation

Production of embryos and subsequent implantation into recipient females to eliminate pathogens from genetically modified (GM) mice. A minimum of two recipient females will be implanted to generate pups.

Genome Editing

Random integration

A highly concentrated aliquot of the plasmid needs to be provided to the GEMF for purification. Ideally, an endotoxin-free MidiPrep or a MaxiPrep should be provided to the GEMF. Restriction enzyme(s) to isolate the linearized transgene also needs to be provided.

Transgenic mice will be generated by pronuclear injection of C57BL/6J zygotes.

Should another background be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

Knock out (KO)

Small or large deletions will be performed in C57BL/6J zygotes.

Should another background be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

Knock in (KI) by electroporation

KI mice will be generated by AAV-driven electroporation of C57BL/6J zygotes using engineered endonucleases.

Should another background be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

Knock in (KI) by pronuclear injection

The donor plasmid used for Homologous Directed Repair (HDR) needs to be provided to the GEMF at high concentration for purification. Ideally, an endotoxin-free MidiPrep or a MaxiPrep should be provided to the GEMF.

KI mice will be generated by pronuclear injection of C57BL/6J zygotes using engineered endonucleases.

Should another background be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

Small genomic modifications

Genetic modifications up to 200 nucleotides will be performed in C57BL/6J zygotes.

Should another background be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

All included — Knock out (KO)

This service includes:

Small or large deletions in C57BL/6 zygotes.
Gel electrophoresis of PCR amplicons (if applicable).
Sanger sequencing or targeted Next Generation Sequencing (NGS) to confirm proper deletion.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

All included — Small genomic modifications

This service includes:

Genetic modifications up to 200 nucleotides in C57BL/6J zygotes.
Sanger sequencing and/or targeted Next Generation Sequencing (NGS) to confirm proper insertion.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

All included — Knock in (KI) by electroporation

This service includes:

AAV-driven electroporation of C57BL/6J zygotes with engineered endonucleases.
Transgene-specific PCR to identify potential Founders
Long Read Sequencing of selected F1 mice (up to three) to confirm proper integration.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

All included — Knock in (KI) by pronuclear injection

This service includes:

Pronuclear injection of C57BL/6J zygotes with engineered endonucleases.
Transgene-specific PCR to identify potential Founders
Long Read Sequencing of selected F1 mice (up to three) to confirm proper integration.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs beyond the weaning date will be charged to the principal investigator.

Molecular Biology

PCR Genotyping

DNA extraction and PCR genotyping to identify potential founders.

Sanger Sequencing

DNA extraction and Sanger sequencing to identify potential founders.

Targeted Next Generation Sequencing (NGS)

DNA extraction and next generation sequencing (NGS) for quality control of the editing events.

Targeted Long Read Sequencing (LRS)

DNA extraction and long read sequencing (LRS) for quality control of the editing events for up to three mice.

Gene Targeting (ES Cells)

For all ES-Cells related projects, the principal investigator will need to provide their competent ES-cell lines. Please contact Fabien Delerue to discuss your project.