Wisconsin National Primate Research Center
Over 60 years of life-saving research and humane animal care
A CRISPR-Cas9 injection into a cell under the microscope

Precision Medicine and Genomic Resources

Critical goals of NHP research include developing genetically precise NHP models for studying the pathogenesis of human genetic diseases, identifying NHP orthologs of human disease-associated alleles, and developing therapeutic strategies for genetic diseases, including regenerative medicine approaches using iPSCs and targeted gene therapies.

This Resource Unit aims to integrate embryonic and cellular genome editing to develop NHP models, advance in vivo cellular targeting and transplantation, and discover “phenotype to genotype” associations by whole genome sequencing analyses. The unique resources developed by the Unit will be available to WNPRC investigators and the broad scientific community.

RESEARCH GOALS

  • Optimize delivery of CRISPR-Cas9 genome editing constructs to NHP cells and embryos.
  • Advance precision MHC-defined NHP models for gene and stem cell-based therapies for AIDS.
  • Expand and formalize Genomics Resources to support investigators in employing genomic data from nonhuman primate models in their research.

Optimizing Cell and Embryo Genome Editing Platforms

CONTACTS

Current Research

  • Targeting CYP19A1 in rhesus macaque cells and embryos to create an aromatase deficiency model. Aromatase deficiency leads to hyperandrogenism, a feature of polycystic ovarian syndrome in humans.
  • Correction of a naturally-occurring mutation, MAPT R406W, in rhesus macaque cells to develop cell-based gene therapies for frontotemporal dementia.

Activities Supported

  • Provide nonhuman primate fibroblasts, embryonic, induced pluripotent, and trophoblast stem cells (ESC, iPSC, and TSC) for testing CRISPR/Cas9 targeting vectors.
  • Guide CRISPR/Cas9 construct design and optimization of in vitro cell cultures
  • Collect oocytes and sperm for in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), providing technical expertise in microinjection and embryo culture.
  • Transfer of in vitro produced embryos to surrogate females and monitor the progress of the pregnancies. Sampling and testing of the offspring for the genetic modification of choice.
  • Tailor project-specific neonatal husbandry to the predicted phenotype of the edited offspring.
  • Develop sperm and embryo cryopreservation for experimental and germplasm banking.

Advancing Precision Therapies

CONTACTS

Current Research

  • Developing MHC-defined model off-the-shelf CAR-T and CAR-NK therapies for HIV infection.
  • Developing non-genotoxic conditioning regimens for transplantation of gene-edited hematopoietic stem cells (HSCs).

Activities Supported

  • Generate and characterize immune effector cells derived from NHP-iPSCs
  • In vivo administration of iPSC-derived hematopoietic progenitors and immune cells
  • Genetic editing of HSCs and iPSCs using CRISPR/Cas9
  • Isolate HSCs from bone marrow and peripheral blood
  • Transplant HSCs and develop novel conditioning regimens
  • Evaluate preclinical iPSC-derived vascular cells

Genomic Discovery

CONTACTS

Current Research

  • Accelerating NHP model development through discovery of naturally-occurring mutations.
  • Analysis of existing and future genomic data to support phenotype-to-genotype and genotype-to-phenotype studies.
  • Comprehensive genomic analysis of CRISPR/Cas9 edited NHP cells and embryos, complementing genome editing and precision therapies research.

Activities Supported

  • Support and assist WNPRC researchers and external investigators in the analysis of genomic data.
  • Provision of sequencing and bioinformatic support for WNPRC researchers and external investigators seeking to perform whole genome or exome sequencing analysis.
  • A goal of Genomic Discovery is to expand and formalize Genomic Resources for investigators to utilize a rapidly increasing NHP genomic database. The PMGR unit offers fee-for-service sequencing and bioinformatic services, in addition to consultation on genomic studies. Investigators seeking genomic support should contact Jeffrey Rogers, Ph.D. Investigators interested specifically in MHC or other immune loci genotyping should contact the WNPRC Genomics Services unit.   

SHARED RESOURCES

A goal of the PMGR unit is to share NHP cell and genomic resources with the broad scientific community.

The following cell resources are available for distribution:

Species PMGR  Unit SCR Unit
mutated wild-type (control) wild-type (control)
Cynomolgus  CCR5-knockout iPSC fibroblasts

trophoblast stem cells

 fibroblasts

iPSCs
Rhesus  MAPT R406W fibroblasts fibroblasts

trophoblast stem cells

 fibroblasts

ESCs iPSCs
Marmoset LRRK2 G2019S ESC

LRRK2 G2019S iPSC

 fibroblasts

ESCs, iPSCs

Highlighted Publications

Peer-Reviewed Publications

Schmidt JK, Reynolds MR, Golos TG, Slukvin II. CRISPR/Cas9 genome editing to create nonhuman primate models for studying stem cell therapies for HIV infection. Retrovirology 2022, 19:17

Weinfurter JT, D’Souza SS, Matschke LM, Bennett S, Kelnhofer-Millevolte LE, Suknuntha K, Kumar A, Coonen J, Capitini CM, Hematti P, Golos TG, Slukvin II, Reynolds MR. Allogeneic MHC-matched T-cell receptor α/β-depleted bone marrow transplants in SHIV-infected, ART-suppressed Mauritian cynomolgus macaques. Sci Rep. 2022 Jul 19;12(1):12345.

D’Souza SS, Kumar A, Maufort J, Weinfurter JT, Raymond M, Strelchenko N, Perrin E, Coonen J, Mejia A, Simmons H, Torbett BE, Reynolds M, Thomson JA, Slukvin I. Assessment of Safety and Immunogenicity of MHC homozygous iPSC-derived CD34+ Hematopoietic Progenitors in a NHP Model. Blood Adv. 2022 Apr 11:bloodadvances.2022006984.

D’Souza SS, Kumar A, Weinfurter J, Park MA, Maufort J, Tao L, Kang H, Dettle ST, Golos T, Thomson JA, Reynolds MR, Slukvin I. Generation of SIV-resistant T cells and macrophages from nonhuman primate induced pluripotent stem cells with edited CCR5 locus. Stem Cell Reports. 2022 Mar 17:S2213-6711(22)00136-9.

Schmidt JK, Jones KM, Van Vleck T, Emborg ME. Modeling genetic diseases in nonhuman primates through embryonic and germline modification: Considerations and challenges. Sci Transl Med. 2022 Mar 2;14(634):eabf4879.

D’Souza SS, Bennett S, Kumar A, Kelnhofer LE, Weinfurter, Suknuntha K, Coonen J, Mejia A, Simmons H, Golos T, Hematti P, Capitini CM, Reynolds MR, Slukvin II. Transplantation of TCRa/b-depleted allogeneic bone marrow in nonhuman primates. Exp. Hematol. S0301-472X(20)30563-4.

Warren WC, Harris RA, Haukness M, Fiddes IT, Murali SC, Fernandes J, Dishuck PC, Storer JM, Raveendran M, Hillier LW, Porubsky D, Mao Y, Gordon D. Vollger MR, Lewis AP, Munson KM, DeVogeleare E, Armstrong J, Diekhans M, Walker JA,  Tomlinson C, Graves-Lindsay TA, Kremitzki M, Salama SR, Audano PA, Escalona M, Maurer NW, Antonacci F, Mercuri L, Maggionlini FAM, Catacchio CR, Underwood JG, O’Connor DH, Sanders AD, Korbel JO, Ferguson B, Kubisch HM, Picker L, Kalin NH, Rosene D, Levine J, Abbott DH, Gray SB, Sanchez MM, Kovacs-Balint ZA, Kemnitz JW, Thomasy SM, Roberts JA, Kinnally EL, Capitanio JP, Skene JHP, Platt M, Cole SA, Green RE, Ventura M, Wiseman RW, Paten B, Batzer MA, Rogers J, Eichler EE. Sequence diversity analyses of an improved rhesus macaque genome enhance its biomedical utility. Science. 2020;370(6523).

Schmidt JK*, Strelchenko N*, Park MA, Kim YH, Mean KD, Schotzko ML, Kang HJ, Golos TG#, Slukvin II#. Genome editing of CCR5 by CRISPR-Cas9 in Mauritian cynomolgus macaque embryos. Sci. Rep. 2020; 10(1):18457.

Strelchenko N, Kropp Schmidt J, Mean KD, Golos TG, Slukvin II. Cryopreservation of Mauritian Cynomolgus Macaque (Macaca fascicularis) Sperm in Chemically Defined Medium. J. Am. Assoc. Lab. Anim. 2020.

Kumar A, D’Souza S, Uenishi G, Park M, Lee J, Slukvin I. Generation of T cells from human and nonhuman primate pluripotent stem cells. Bio-protocol 2020; 10:e.3675.

Vermilyea SC, Babinski A, Tran N, To S, Guthrie S, Kluss JH, Schmidt JK, Wiepz GJ, Meyer MG, Murphy ME, Cookson MR, Emborg ME. In Vitro CRISPR/Cas9-Directed Gene Editing to Model LRRK2 G2019S Parkinson’s Disease in Common Marmosets. Sci. Rep. 2020; 10:3447.

Brok-Volchanskaya VS, Bennin DA, Suknuntha K, Klemm LC, Huttenlocher A, Slukvin I. Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA. Stem Cell Rep. 2019; 13.

Maufort JP, Israel JS, Brown ME, Kempton SJ, Albano NJ, Zen WF, Kelnhofer LE, Reynolds MR, Perrin ES, Sanchez RJ, Sluvkin, II, Thomson JA, Poore So. Major Histocompatibility Complex-Matched Arteries Have Similar Patency to Autologous Arteries in a Mauritian Cynomolgus Macaque Major Histocompatibility Complex-Defined Transplant Model. J. Am. Heart Assoc. 2019; 8.

Vermilyea SC, Guthrie S, Meyer M, Smuga-Otto K, Braun K, Howden S, Thomson JA, Zhang SC, Emborg ME, Golos TG. Induced Pluripotent Stem Cell-Derived Dopaminergic Neurons from Adult Common Marmoset Fibroblasts. Stem Cells Dev 2017; 26:1225-1235.