Molecular genetic tools have had a profound impact on neuroscience, but until recently their application has largely been confined to a few model species, most notably mouse, zebrafish, Drosophila melanogaster and Caenorhabditis elegans. With the development of new genome engineering technologies such as CRISPR, it is becoming increasingly feasible to apply these molecular tools in a wider range of species, including nonhuman primates. This will lead to many opportunities for brain research, but it will also pose challenges. Here we identify some of these opportunities and challenges in light of recent and foreseeable technological advances and offer some suggestions. Our main focus is on the creation of new primate disease models for understanding the pathological mechanisms of brain disorders and for developing new approaches to effective treatment. However, we also emphasize that primate genetic models have great potential to address many fundamental questions about brain function, providing an essential foundation for future progress in disease research.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Murray, C.J.L. et al. Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990-2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380, 2197–2223 (2012).
Roehrig, C. Mental disorders top the list of the most costly conditions in the United States: $201 billion. Health Aff. (Millwood) 35, 1130–1135 (2016).
Hyman, S.E. Revolution stalled. Sci. Transl. Med. 4, 155cm11 (2012).
Gribkoff, V.K. & Kaczmarek, L.K. The need for new approaches in CNS drug discovery: why drugs have failed, and what can be done to improve outcomes. Neuropharmacology http://dx.doi.org/10.1016/j.neuropharm.2016.03.021 (2016).
Kaitin, K.I. & DiMasi, J.A. Pharmaceutical innovation in the 21st century: new drug approvals in the first decade, 2000–2009. Clin. Pharmacol. Ther. 89, 183–188 (2011).
Amplion. Clinical Development Success Rates 2006–2015 http://www.amplion.com/clinical-development-success-rates (2016).
Fidler, B. After failed schizophrenia trial, Forum Pharma to shutter this week. Xconomy http://www.xconomy.com/boston/2016/06/27/after-failed-schizophrenia-trial-forum-pharma-to-shutter-this-week/ 2016).
Pangalos, M.N., Schechter, L.E. & Hurko, O. Drug development for CNS disorders: strategies for balancing risk and reducing attrition. Nat. Rev. Drug Discov. 6, 521–532 (2007).
Nestler, E.J. & Hyman, S.E. Animal models of neuropsychiatric disorders. Nat. Neurosci. 13, 1161–1169 (2010).
Fernando, A.B.P. & Robbins, T.W. Animal models of neuropsychiatric disorders. Annu. Rev. Clin. Psychol. 7, 39–61 (2011).
Sarter, M. & Tricklebank, M. Revitalizing psychiatric drug discovery. Nat. Rev. Drug Discov. 11, 423–424 (2012).
Institute of Medicine (US) Forum on Neuroscience and Nervous System Disorders. Improving the Utility and Translation of Animal Models for Nervous System Disorders: Workshop Summary (National Academies Press, 2013).
Bluemel, J., Korte, S., Schenck, E. & Weinbauer, G. (eds.) The Nonhuman Primate in Nonclinical Drug Development and Safety Assessment 1st edn. (Elsevier, 2016).
Chan, A.W., Chong, K.Y., Martinovich, C., Simerly, C. & Schatten, G. Transgenic monkeys produced by retroviral gene transfer into mature oocytes. Science 291, 309–312 (2001).
Yang, S.-H. et al. Towards a transgenic model of Huntington's disease in a non-human primate. Nature 453, 921–924 (2008).
Sasaki, E. et al. Generation of transgenic non-human primates with germline transmission. Nature 459, 523–527 (2009).
Chan, A.W.S. Progress and prospects for genetic modification of nonhuman primate models in biomedical research. ILAR J. 54, 211–223 (2013).
Okano, H., Miyawaki, A. & Kasai, K. Brain/MINDS: brain-mapping project in Japan. Phil. Trans. R. Soc. Lond. B http://dx.doi.org/10.1098/rstb.2014.0310 (2015).
Cyranoski, D. Monkey kingdom. Nature 532, 300–302 (2016).
Miller, J.C. et al. A TALE nuclease architecture for efficient genome editing. Nat. Biotechnol. 29, 143–148 (2011).
Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013).
Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823–826 (2013).
Carlson, D.F. et al. Efficient TALEN-mediated gene knockout in livestock. Proc. Natl. Acad. Sci. USA 109, 17382–17387 (2012).
Wang, H. et al. One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153, 910–918 (2013).
Niu, Y. et al. Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos. Cell 156, 836–843 (2014).
Liu, H. et al. TALEN-mediated gene mutagenesis in rhesus and cynomolgus monkeys. Cell Stem Cell 14, 323–328 (2014).
Chen, Y. et al. Functional disruption of the dystrophin gene in rhesus monkey using CRISPR/Cas9. Hum. Mol. Genet. 24, 3764–3774 (2015).
Sasaki, E. Prospects for genetically modified non-human primate models, including the common marmoset. Neurosci. Res. 93, 110–115 (2015).
Whitelaw, C.B.A., Sheets, T.P., Lillico, S.G. & Telugu, B.P. Engineering large animal models of human disease. J. Pathol. 238, 247–256 (2016).
Liang, P. et al. CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes. Protein Cell 6, 363–372 (2015).
Willsey, A.J. & State, M.W. Autism spectrum disorders: from genes to neurobiology. Curr. Opin. Neurobiol. 30, 92–99 (2015).
Neale, B.M. & Sklar, P. Genetic analysis of schizophrenia and bipolar disorder reveals polygenicity but also suggests new directions for molecular interrogation. Curr. Opin. Neurobiol. 30, 131–138 (2015).
Del-Aguila, J.L. et al. Alzheimer's disease: rare variants with large effect sizes. Curr. Opin. Genet. Dev. 33, 49–55 (2015).
Watakabe, A. et al. Comparative analyses of adeno-associated viral vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral cortex. Neurosci. Res. 93, 144–157 (2015).
Deverman, B.E. et al. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nat. Biotechnol. 34, 204–209 (2016).
Choudhury, S.R. et al. Viral vectors for therapy of neurologic diseases. Neuropharmacology http://dx.doi.org/10.1016/j.neuropharm.2016.02.013 (2016).
Han, X. et al. Millisecond-timescale optical control of neural dynamics in the nonhuman primate brain. Neuron 62, 191–198 (2009).
Diester, I. et al. An optogenetic toolbox designed for primates. Nat. Neurosci. 14, 387–397 (2011).
Macosko, E.Z. et al. Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161, 1202–1214 (2015).
Tasic, B. et al. Adult mouse cortical cell taxonomy revealed by single cell transcriptomics. Nat. Neurosci. 19, 335–346 (2016).
Krishnaswami, S.R. et al. Using single nuclei for RNA-seq to capture the transcriptome of postmortem neurons. Nat. Protoc. 11, 499–524 (2016).
Anderson, D.J. & Perona, P. Toward a science of computational ethology. Neuron 84, 18–31 (2014).
Sadagopan, S., Temiz-Karayol, N.Z. & Voss, H.U. High-field functional magnetic resonance imaging of vocalization processing in marmosets. Sci. Rep. 5, 10950 (2015).
Herrmann, T. et al. The travelling-wave primate system: a new solution for magnetic resonance imaging of macaque monkeys at 7 Tesla ultra-high field. PLoS One 10, e0129371 (2015).
Eliades, S.J. & Wang, X. Chronic multi-electrode neural recording in free-roaming monkeys. J. Neurosci. Methods 172, 201–214 (2008).
Roy, S. & Wang, X. Wireless multi-channel single unit recording in freely moving and vocalizing primates. J. Neurosci. Methods 203, 28–40 (2012).
Yin, M. et al. Wireless neurosensor for full-spectrum electrophysiology recordings during free behavior. Neuron 84, 1170–1182 (2014).
Fernandez-Leon, J.A. et al. A wireless transmission neural interface system for unconstrained non-human primates. J. Neural Eng. 12, 056005 (2015).
Izpisua Belmonte, J.C. et al. Brains, genes, and primates. Neuron 86, 617–631 (2015).
Vallender, E.J. & Miller, G.M. Nonhuman primate models in the genomic era: a paradigm shift. ILAR J. 54, 154–165 (2013).
Kaiser, T. & Feng, G. Modeling psychiatric disorders for developing effective treatments. Nat. Med. 21, 979–988 (2015).
Tu, Z., Yang, W., Yan, S., Guo, X. & Li, X.-J. CRISPR/Cas9:a powerful genetic engineering tool for establishing large animal models of neurodegenerative diseases. Mol. Neurodegener. 10, 35 (2015).
Okano, H., Hikishima, K., Iriki, A. & Sasaki, E. The common marmoset as a novel animal model system for biomedical and neuroscience research applications. Semin. Fetal Neonatal Med. 17, 336–340 (2012).
Bendor, D. & Wang, X. The neuronal representation of pitch in primate auditory cortex. Nature 436, 1161–1165 (2005).
Song, X., Osmanski, M.S., Guo, Y. & Wang, X. Complex pitch perception mechanisms are shared by humans and a New World monkey. Proc. Natl. Acad. Sci. USA 113, 781–786 (2016).
Mitchell, J.F., Reynolds, J.H. & Miller, C.T. Active vision in marmosets: a model system for visual neuroscience. J. Neurosci. 34, 1183–1194 (2014).
Shiba, Y., Santangelo, A.M. & Roberts, A.C. Beyond the medial regions of prefrontal cortex in the regulation of fear and anxiety. Front. Syst. Neurosci. 10, 12 (2016).
Miller, C.T. et al. Marmosets: a neuroscientific model of human social behavior. Neuron 90, 219–233 (2016).
Kaas, J.H. The evolution of brains from early mammals to humans. Wiley Interdiscip. Rev. Cogn. Sci. 4, 33–45 (2013).
Mashiko, H. et al. Comparative anatomy of marmoset and mouse cortex from genomic expression. J. Neurosci. 32, 5039–5053 (2012).
Liu, D. et al. Medial prefrontal activity during delay period contributes to learning of a working memory task. Science 346, 458–463 (2014).
Leach, P.T., Hayes, J., Pride, M., Silverman, J.L. & Crawley, J.N. Normal performance of Fmr1 mice on a touchscreen delayed nonmatching to position working memory task. eNeuro http://dx.doi.org/10.1523/eneuro.0143-15.2016 (2016).
Shultz, S., Opie, C. & Atkinson, Q.D. Stepwise evolution of stable sociality in primates. Nature 479, 219–222 (2011).
Donaldson, Z.R. & Young, L.J. Oxytocin, vasopressin, and the neurogenetics of sociality. Science 322, 900–904 (2008).
Garrison, J.L. et al. Oxytocin/vasopressin-related peptides have an ancient role in reproductive behavior. Science 338, 540–543 (2012).
Freeman, S.M. & Young, L.J. Comparative perspectives on oxytocin and vasopressin receptor research in rodents and primates: translational implications. J. Neuroendocrinol. http://dx.doi.org/10.1111/jne.12382 (2016).
Levin, E.D. α7-Nicotinic receptors and cognition. Curr. Drug Targets 13, 602–606 (2012).
Quik, M. et al. Localization of nicotinic receptor subunit mRNAs in monkey brain by in situ hybridization. J. Comp. Neurol. 425, 58–69 (2000).
Fidler, B. Restructuring looms for Forum as neuro drug fails key clinical test. Xconomy http://www.xconomy.com/boston/2016/03/24/restructuring-looms-for-forum-as-neuro-drug-fails-key-clinical-test/ (2016).
Lemon, R.N. Descending pathways in motor control. Annu. Rev. Neurosci. 31, 195–218 (2008).
Butti, C., Santos, M., Uppal, N. & Hof, P.R. Von Economo neurons: clinical and evolutionary perspectives. Cortex 49, 312–326 (2013).
Herculano-Houzel, S., Collins, C.E., Wong, P. & Kaas, J.H. Cellular scaling rules for primate brains. Proc. Natl. Acad. Sci. USA 104, 3562–3567 (2007).
Ventura-Antunes, L., Mota, B. & Herculano-Houzel, S. Different scaling of white matter volume, cortical connectivity, and gyrification across rodent and primate brains. Front. Neuroanat. 7, 3 (2013).
Takeda, S. et al. Neuronal uptake and propagation of a rare phosphorylated high-molecular-weight tau derived from Alzheimer's disease brain. Nat. Commun. 6, 8490 (2015).
LaFerla, F.M. & Green, K.N. Animal models of Alzheimer disease. Cold Spring Harb. Perspect. Med. 2, a006320 (2012).
Cummings, J.L., Morstorf, T. & Zhong, K. Alzheimer's disease drug-development pipeline: few candidates, frequent failures. Alzheimers Res. Ther. 6, 37 (2014).
Forny-Germano, L. et al. Alzheimer's disease-like pathology induced by amyloid-β oligomers in nonhuman primates. J. Neurosci. 34, 13629–13643 (2014).
Johnson, M.B. et al. Single-cell analysis reveals transcriptional heterogeneity of neural progenitors in human cortex. Nat. Neurosci. 18, 637–646 (2015).
Otani, T., Marchetto, M.C., Gage, F.H., Simons, B.D. & Livesey, F.J. 2D and 3D stem cell models of primate cortical development identify species-specific differences in progenitor behavior contributing to brain size. Cell Stem Cell 18, 467–480 (2016).
Webb, S.J., Jones, E.J.H., Kelly, J. & Dawson, G. The motivation for very early intervention for infants at high risk for autism spectrum disorders. Int. J. Speech Lang Pathol. 16, 36–42 (2014).
Qureshi, I.A. & Mehler, M.F. Emerging roles of non-coding RNAs in brain evolution, development, plasticity and disease. Nat. Rev. Neurosci. 13, 528–541 (2012).
Walker, S. & Scherer, S.W. Identification of candidate intergenic risk loci in autism spectrum disorder. BMC Genomics 14, 499 (2013).
Batzoglou, S., Pachter, L., Mesirov, J.P., Berger, B. & Lander, E.S. Human and mouse gene structure: comparative analysis and application to exon prediction. Genome Res. 10, 950–958 (2000).
Bernard, A. et al. Transcriptional architecture of the primate neocortex. Neuron 73, 1083–1099 (2012).
Bateson, P. Review of Research using Non-Human Primates http://www.bbsrc.ac.uk/documents/review-research-using-nhps-pdf/ (2011).
Emanuel, E.J., Wendler, D. & Grady, C. What makes clinical research ethical? J. Am. Med. Assoc. 283, 2701–2711 (2000).
Liu, Z. et al. Autism-like behaviours and germline transmission in transgenic monkeys overexpressing MeCP2. Nature 530, 98–102 (2016).
Liu, Z. et al. Generation of macaques with sperm derived from juvenile monkey testicular xenografts. Cell Res. 26, 139–142 (2016).
Kanatsu-Shinohara, M. et al. Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Biol. Reprod. 69, 612–616 (2003).
Sato, T. et al. Genome editing in mouse spermatogonial stem cell lines using TALEN and double-nicking CRISPR/Cas9. Stem Cell Reports 5, 75–82 (2015).
Zhou, Q. et al. Complete meiosis from embryonic stem cell-derived germ cells in vitro. Cell Stem Cell 18, 330–340 (2016).
Campbell, K.H., McWhir, J., Ritchie, W.A. & Wilmut, I. Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–66 (1996).
Hrabeěde Angelis, M. et al. Analysis of mammalian gene function through broad-based phenotypic screens across a consortium of mouse clinics. Nat. Genet. 47, 969–978 (2015).
Perrin, S. Preclinical research: make mouse studies work. Nature 507, 423–425 (2014).
Berry-Kravis, E.M. et al. Effects of STX209 (arbaclofen) on neurobehavioral function in children and adults with fragile X syndrome: a randomized, controlled, phase 2 trial. Sci. Transl. Med. 4, 152ra127 (2012).
Berry-Kravis, E. et al. Mavoglurant in Fragile X syndrome: results of two randomized, double-blind, placebo-controlled trials. Sci. Transl. Med. 8, 321ra5 (2016).
Lombardi, L.M., Baker, S.A. & Zoghbi, H.Y. MECP2 disorders: from the clinic to mice and back. J. Clin. Invest. 125, 2914–2923 (2015).
Bourgeron, T. Current knowledge on the genetics of autism and propositions for future research. C. R. Biol. 339, 300–307 (2016).
Agamaite, J.A., Chang, C.-J., Osmanski, M.S. & Wang, X. A quantitative acoustic analysis of the vocal repertoire of the common marmoset (Callithrix jacchus). J. Acoust. Soc. Am. 138, 2906–2928 (2015).
Clarke, H.F., Horst, N.K. & Roberts, A.C. Regional inactivations of primate ventral prefrontal cortex reveal two distinct mechanisms underlying negative bias in decision making. Proc. Natl. Acad. Sci. USA 112, 4176–4181 (2015).
Kangas, B.D., Bergman, J. & Coyle, J.T. Touchscreen assays of learning, response inhibition, and motivation in the marmoset (Callithrix jacchus). Anim. Cogn. 19, 673–677 (2016).
Ioannidis, J.P.A. Why most published research findings are false. PLoS Med. 2, e124 (2005).
Collins, F.S. & Tabak, L.A. Policy: NIH plans to enhance reproducibility. Nature 505, 612–613 (2014).
Button, K.S. et al. Power failure: why small sample size undermines the reliability of neuroscience. Nat. Rev. Neurosci. 14, 365–376 (2013).
Open Science Collaboration. Psychology. Estimating the reproducibility of psychological science. Science 349, aac4716 (2015).
Tsilidis, K.K. et al. Evaluation of excess significance bias in animal studies of neurological diseases. PLoS Biol. 11, e1001609 (2013).
Bauer, S.A. & Baker, K.C. Persistent effects of peer rearing on abnormal and species-appropriate activities but not social behavior in group-housed rhesus macaques (Macaca mulatta). Comp. Med. 66, 129–136 (2016).
Committee on Strategies for Small-Number-Participant Clinical Research Trials, Board on Health Sciences Policy & Institute of Medicine. Small Clinical Trials: Issues and Challenges (National Academies Press, 2001).
National Research Council (US) Institute for Laboratory Animal Research. Transportation of Primates and the Animal Welfare Act (National Academies Press, 2003).
Bloom, D.E. et al. The Global Economic Burden of Noncommunicable Diseases (World Economic Forum, Geneva, 2011).
Chisholm, D. et al. Scaling-up treatment of depression and anxiety: a global return on investment analysis. Lancet Psychiatry 3, 415–424 (2016).
Patel, V. et al. Addressing the burden of mental, neurological, and substance use disorders: key messages from Disease Control Priorities, 3rd edition. Lancet 387, 1672–1685 (2016).
Chan, A.W.S. et al. Progressive cognitive deficit, motor impairment and striatal pathology in a transgenic Huntington disease monkey model from infancy to adulthood. PLoS One 10, e0122335 (2015).
Niu, Y. et al. Early Parkinson's disease symptoms in α-synuclein transgenic monkeys. Hum. Mol. Genet. 24, 2308–2317 (2015).
Cox, D.B.T., Platt, R.J. & Zhang, F. Therapeutic genome editing: prospects and challenges. Nat. Med. 21, 121–131 (2015).
Yang, L. et al. Genome-wide inactivation of porcine endogenous retroviruses (PERVs). Science 350, 1101–1104 (2015).
Slaymaker, I.M. et al. Rationally engineered Cas9 nucleases with improved specificity. Science 351, 84–88 (2016).
Kleinstiver, B.P. et al. High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature 529, 490–495 (2016).
Wan, H. et al. One-step generation of p53 gene biallelic mutant cynomolgus monkey via the CRISPR/Cas system. Cell Res. 25, 258–261 (2015).
Chen, Y. et al. Generation of cynomolgus monkey chimeric fetuses using embryonic stem cells. Cell Stem Cell 17, 116–124 (2015).
Swiech, L. et al. In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9. Nat. Biotechnol. 33, 102–106 (2015).
Platt, R.J. et al. CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell 159, 440–455 (2014).
Konermann, S. et al. Optical control of mammalian endogenous transcription and epigenetic states. Nature 500, 472–476 (2013).
Konermann, S. et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517, 583–588 (2015).
Dominguez, A.A., Lim, W.A. & Qi, L.S. Beyond editing: repurposing CRISPR-Cas9 for precision genome regulation and interrogation. Nat. Rev. Mol. Cell Biol. 17, 5–15 (2016).
Komor, A.C., Kim, Y.B., Packer, M.S., Zuris, J.A. & Liu, D.R. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 533, 420–424 (2016).
Abudayyeh, O.O. et al. C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science http://dx.doi.org/10.1126/science.aaf5573 (2016).
Research projects related to this work at MIT and Broad Institute are supported by the Poitras Center for Affective Disorders Research, the Stanley Center for Psychiatric Disorders Research at Broad Institute of MIT and Harvard, CHDI, Global Academic Innovation Partnering at F. Hoffmann-La Roche Ltd, The Brain Research Foundation, the Massachusetts Life Sciences Center, NIH BRAIN Initiative and Edward and Kay Poitras. Related research at Shenzhen Institutes of Advanced Technology is supported by a Shenzhen Peacock plan grant (KQTD20140630180249366) and by the Guangdong Innovative and Entrepreneurial Research Team Program (No. 2014ZT05S020). L.W. is supported by the CAS Strategic Priority Research Program (XDB02050003) and National Science Fund for Distinguished Young Scholars (No. 81425010). H.Z. is supported by a Shenzhen Subject Arrangement Basic Research Grant (JCYJ20151030140325151) and by the CAS Hundred Talent program. Some of the ideas presented here emerged from discussions at a symposium on transgenic primate research that was held at Shenzhen Institutes of Advanced Technology in China on March 22–23, 2016. We thank all the participants at that meeting for their contributions, and we thank the Ministry of Science and Technology of China and the Chinese Academy of Sciences for financial support of the meeting.
The authors declare no competing financial interests.
About this article
Deep convolutional network for animal sound classification and source attribution using dual audio recordings
The Journal of the Acoustical Society of America (2019)
National Science Review (2019)
Transgenic rhesus monkeys carrying the human MCPH1 gene copies show human-like neoteny of brain development
National Science Review (2019)
Animal Models and Experimental Medicine (2019)
International Journal of Experimental Pathology (2019)