Mouse Engineering Core

The Mouse Engineering Core was established in 1992 operating at the BWH, and is now located at Harvard Medical School. The Mouse Engineering Core provides services for the generation of transgenic and knockout mice using state-of-the-art facilities and equipment.

The Core has all of the equipment necessary for generation of transgenic mice including a Nikon Diaphot microscope equipped with Nomarski Optics and Narishige micromanipulators for microinjections, Nikon surgical microscopes for egg isolation and transfer, a sutter needle puller and a de Fonbrune microforge. The facility maintains mice necessary for egg donors, egg recipients, and vasectomized males.

ES cell culture services have been offered for 15 years. The Core offers electroporation of targeting vectors into ES cells and provides investigators with DNA to identify ES cells carrying the desired recombination events; and ES cell expansion for microinjection into blastocysts.

The Core's newest services include CRISPR injections, sperm and embryo cryopreservation, and reanimation with fresh or frozen sperm via IVF or the thawing and implanting embryos.

Publication Acknowledgement

If research supported by this core facility results in publication, please acknowledge this support by including the following in your publication(s):

We thank Harvard Cancer Consortium in Boston, MA, for the use of the Mouse Engineering Core, which provided ______________________________ service. Harvard Cancer Consortium is supported in part by a NCI Cancer Center Support Grant # NIH 5 P30 CA06516.

• CRISPR Injection services
• ES cell electroporation services
• ES cell injection services
• Fresh/Frozen sperm IVF
• Inducible progenitor stem cells (IPS) injection
• Injection of DNA constructs (BALB C, C57BL/6, FVB) into egg pronuclei for generating transgenic mice
• Lentiviral Injection services
• Sperm/embryo cryopreservation
• Tail biopsy services
• Thawing and implantation of embryos

Baseline expression of c-Myc defines the tissue specificity of oncogenic K-Ras

Principal Investigators: Kevin M Haigis DFCI (Cancer Genetics and Epigenetics and Gastrointestinal Malignancies programs)

Description: KRAS is among the most frequently mutated oncogenes in cancer. Yet, mutations in KRAS are common only in tumors originating from a subset of tissues. It is critical to understand the molecular mechanisms underlying this oncogene tissue specificity. Utilizing genetically engineered mouse models carrying a conditional oncogenic allele of K-ras , we expressed activated K-Ras in adult tissues to investigate its specificity. We discovered that the ability of K-Ras G12D to influence the fitness of cells in a given tissue is not determined by its canonical signaling through MAPK. Instead, low baseline expression of c-Myc renders tissues non-permissive to oncogenic K-Ras, a context that can be reversed in the liver by ectopically expressing c-Myc. This functions independently of the proliferative index of the tissue or the induction of cell cycle arrest or apoptosis. Our findings reveal the importance of the basal state of the tissue-inherent signaling network for determining oncogene specificity. ​

Contribution of the Core: KrasLSL-WT mice were generated by blastocyst injections using KrasLGL-WT ES cells
that were a gift from David Tuveson.​

Publication

Olesja Popow, Qing Yu, Clarance Yapp, Shannon Hull, João A Paulo, Benjamin Hanna, Shidong Xu, Yanan Kuang, Carlie Sigel, Cloud P Paweletz, Steven P Gygi, Kevin M Haigis. bioRxiv[Preprint]. 2025 Jul 25:2025.07.21.665965. PMID 40777332.​​

Transcription factor TCF1 binds to RORγt and orchestrates a regulatory network that determines homeostatic Th17 cell state

Principal Investigators: Ana C Anderson BWH (Cancer Immunology Program at BWH)

Description: Th17 cells encompass a spectrum of cell states including cells that maintain homeostatic tissue functions and pro-inflammatory cells that can drive autoimmune tissue damage. Identifying regulators that determine Th17 cell states can identify ways to control tissue inflammation and restore homeostasis. Here, we found that IL-23, a cytokine critical for inducing pro-inflammatory Th17 cells, decreased transcription factor TCF1 expression. Conditional deletion of TCF1 in mature T cells increased the pro-inflammatory potential of Th17 cells even in the absence of IL-23 receptor signaling and conferred pro-inflammatory potential to homeostatic Th17 cells. Conversely, sustained TCF1 expression decreased pro-inflammatory Th17 potential. Mechanistically, TCF1 bound to RORγt, thereby interfering with its pro-inflammatory functions, and orchestrated a regulatory network that determined Th17 cell state. Our findings identify TCF1 as a major determinant of Th17 cell state and provide important insight for the development of therapies for Th17-driven inflammatory diseases.​

Contribution of the Core: Generation of CD2-Tcf7L-Tg mice- construct injected in B6 oocytes​

Publication

Davide Mangani, Ayshwarya Subramanian, Linglin Huang, Hanning Cheng, S Harsha Krovi, Yufan Wu, Dandan Yang, Thais G
Moreira, Giulia Escobar, Alexandra Schnell, Karen O Dixon, Rajesh K Krishnan, Vasundhara Singh, Raymond A Sobel, Howard L Weiner, Vijay K Kuchroo, Ana C Anderson. Immunity. 2024 Oct 23;57(11):2565–2582. PMID: 39447575.​

Targeting PGLYRP1 promotes antitumor immunity while inhibiting autoimmune neuroinflammation

Principal Investigators: Vijay K Kuchroo BWH Cancer Immunology Program

Description: Co-inhibitory and checkpoint molecules suppress T cell function in the tumor microenvironment, thereby rendering T cells dysfunctional. Although immune checkpoint blockade is a successful treatment option for multiple human cancers, severe autoimmune-like adverse effects can limit its application. Here, we show that the gene encoding peptidoglycan recognition protein 1 (PGLYRP1) is highly coexpressed with genes encoding co-inhibitory molecules, indicating that it might be a promising target for cancer immunotherapy. Genetic deletion of Pglyrp1 in mice led to decreased tumor growth and an increased activation/effector phenotype in CD8+ T cells, suggesting an inhibitory function of PGLYRP1 in CD8+ T cells. Surprisingly, genetic deletion of Pglyrp1 protected against the development of experimental autoimmune encephalomyelitis, a model of autoimmune disease in the central nervous system. PGLYRP1-deficient myeloid cells had a defect in antigen presentation and T cell activation, indicating that PGLYRP1 might function as a proinflammatory molecule in myeloid cells during autoimmunity. These results highlight PGLYRP1 as a promising target for immunotherapy that, when targeted, elicits a potent antitumor immune response while protecting against some forms of tissue inflammation and autoimmunity. ​

Contribution of the Core: The Core expanded an ES cell line obtained from KOMP and microinjected the EScells into blastocysts​.​

Publication

Alexandra Schnell, Linglin Huang, Brianna M L Regan, Vasundhara Singh, Dominik Vonficht, Alina Bollhagen, Mona Wang, Yu Hou, Lloyd Bod, Raymond A Sobel, Norio Chihara, Asaf Madi, Ana C Anderson, Aviv Regev, Vijay K Kuchroo. Nat Immunol. 2023 Nov;24(11):1908-1920. PMID 37828379.​​​