An enhancer-AAV toolbox to target and manipulate distinct interneuron subtypes

Elisabetta Furlanis¹, Min Dai¹, Brenda Leyva Garcia¹, Thien Tran¹, Josselyn Vergara¹, Ana Pereira¹, Bram L Gorissen¹, Sara Wills¹, Anna Vlachos², Ariel Hairston³, Deepanjali Dwivedi¹, Sarah Du¹, Justin McMahon¹, Shuhan Huang³, Annunziato Morabito³, Arenski Vazquez⁴, Soyoun Kim⁴, Anthony T Lee⁵, Edward F Chang⁵, Taha Razzaq⁶, Ahmed Qazi⁶, Geoffrey Vargish², Xiaoqing Yuan², Adam Caccavano², Steven Hunt², Ramesh Chittajallu², Nadiya McLean², Lauren Hewitt², Emily Paranzino², Haley Rice², Alex C Cummins⁷, Anya Plotnikova⁷, Arya Mohanty⁷, Anne Claire Tangen⁷, Jung Hoon Shin⁷, Reza Azadi⁷, Mark A G Eldridge⁷, Veronica A Alvarez⁷, Bruno B Averbeck⁷, Mansour Alyahyay⁸, Tania Reyes Vallejo⁸, Mohammed Soheib⁸, Lucas G Vattino⁹, Cathryn P MacGregor⁹, Carolina Piletti Chatain¹⁰, Emmie Banks¹¹, Viktor Janos Olah¹¹, Shovan Naskar¹², Sophie Hill¹³, Sophie Liebergall¹³, Rohan Badiani¹, Lili Hyde¹, Ella Hanley¹, Qing Xu¹⁴, Kathryn C Allaway¹, Ethan M Goldberg¹³, Matthew J M Rowan¹¹, Tomasz J Nowakowski⁵, Soohyun Lee¹², Emilia Favuzzi¹⁰, Pascal S Kaeser³, Lucas Sjulson¹⁵, Renata Batista-Brito¹⁵, Anne E Takesian⁹, Leena A Ibrahim⁸, Asim Iqbal⁶, Kenneth A Pelkey², Chris J McBain², Jordane Dimidschstein¹⁶, Gord Fishell¹⁷, Yating Wang¹⁸

Affiliations

¹ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
² Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD 20892, USA.
³ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA.
⁴ Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY, USA.
⁵ Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA, USA; Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA, USA.
⁶ Tibbling Technologies, Redmond, WA, USA.
⁷ National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892-4415, USA.
⁸ Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia.
⁹ Eaton-Peabody Laboratories, Massachusetts Eye and Ear Infirmary, Boston, MA, USA; Department of Otolaryngology - Head and Neck Surgery, Harvard Medical School, Boston, MA, USA.
¹⁰ Department of Neuroscience, Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, CT 06510, USA; Wu Tsai Institute, Yale University, New Haven, CT 06510, USA.
¹¹ Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
¹² Unit of Functional Neural Circuit, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
¹³ Department of Neuroscience, The University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
¹⁴ Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia; Genomics & Systems Biology, New York University Abu Dhabi, Abu Dhabi, UAE.
¹⁵ Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY, USA; Dominick P. Purpura Department of Psychiatry and Behavioral Sciences, Albert Einstein College of Medicine, Bronx, NY, USA.
¹⁶ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: jordane.dimidschstein@gmail.com.
¹⁷ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: gfishell@broadinstitute.org.
¹⁸ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: yawang@broadinstitute.org.

PMID: 40403705
DOI: 10.1016/j.neuron.2025.05.002

An enhancer-AAV toolbox to target and manipulate distinct interneuron subtypes

Elisabetta Furlanis et al. Neuron. 2025.

. 2025 May 21;113(10):1525-1547.e15.

doi: 10.1016/j.neuron.2025.05.002.

Authors

Affiliations

¹ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
² Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD 20892, USA.
³ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA.
⁴ Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY, USA.
⁵ Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA, USA; Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA, USA.
⁶ Tibbling Technologies, Redmond, WA, USA.
⁷ National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892-4415, USA.
⁸ Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia.
⁹ Eaton-Peabody Laboratories, Massachusetts Eye and Ear Infirmary, Boston, MA, USA; Department of Otolaryngology - Head and Neck Surgery, Harvard Medical School, Boston, MA, USA.
¹⁰ Department of Neuroscience, Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, CT 06510, USA; Wu Tsai Institute, Yale University, New Haven, CT 06510, USA.
¹¹ Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
¹² Unit of Functional Neural Circuit, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
¹³ Department of Neuroscience, The University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
¹⁴ Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia; Genomics & Systems Biology, New York University Abu Dhabi, Abu Dhabi, UAE.
¹⁵ Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY, USA; Dominick P. Purpura Department of Psychiatry and Behavioral Sciences, Albert Einstein College of Medicine, Bronx, NY, USA.
¹⁶ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: jordane.dimidschstein@gmail.com.
¹⁷ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: gfishell@broadinstitute.org.
¹⁸ Harvard Medical School, Blavatnik Institute, Department of Neurobiology, Boston, MA 02115, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: yawang@broadinstitute.org.

PMID: 40403705
DOI: 10.1016/j.neuron.2025.05.002

Abstract

In recent years, we and others have identified a number of enhancers that, when incorporated into rAAV vectors, can restrict the transgene expression to particular neuronal populations. Yet, viral tools to access and manipulate specific neuronal subtypes are still limited. Here, we performed systematic analysis of single-cell genomic data to identify enhancer candidates for each of the telencephalic interneuron subtypes. We established a set of enhancer-AAV tools that are highly specific for distinct cortical interneuron populations and striatal cholinergic interneurons. These enhancers, when used in the context of different effectors, can target (fluorescent proteins), observe activity (GCaMP), and manipulate (opto-genetics) specific neuronal subtypes. We also validated our enhancer-AAV tools across species. Thus, we provide the field with a powerful set of tools to study neural circuits and functions and to develop precise and targeted therapy.

Keywords: GABAergic; adeno-associated virus; cholinergic; cortex; enhancer; interneuron.

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Conflict of interest statement

Declaration of interests G.F. is a founder of Regel Therapeutics, which has no competing interests with the present manuscript. G.F. is an advisor for Neuron and Annual Review of Neuroscience. J.D. and K.C.A. are employees of Regel Therapeutics, and J.D. is also a founder. A.I. is the founder of Tibbling Technologies. Patents are pending on all enhancers present in this manuscript. G.F. and J.D. hold the patent for BiCHATe27 and BiSSTe10. The remainder of enhancer patents are held by G.F., M.D., and Y.W.

Update of

An enhancer-AAV toolbox to target and manipulate distinct interneuron subtypes.
Furlanis E, Dai M, Garcia BL, Tran T, Vergara J, Pereira A, Gorissen BL, Sara Wills, Vlachos A, Hairston A, Dwivedi D, Du S, McMahon J, Huang S, Morabito A, Vazquez A, Kim S, Lee AT, Chang EF, Razzaq T, Qazi A, Vargish G, Yuan X, Caccavano A, Hunt S, Chittajallu R, McLean N, Hewitt L, Paranzino E, Rice H, Cummins AC, Plotnikova A, Mohanty A, Tangen AC, Shin JH, Azadi R, Eldridge MAG, Alvarez VA, Averbeck BB, Alyahyay M, Vallejo TR, Soheib M, Vattino LG, MacGregor CP, Chatain CP, Banks E, Olah VJ, Naskar S, Hill S, Liebergall S, Badiani R, Hyde L, Hanley E, Xu Q, Allaway KC, Goldberg EM, Rowan MJM, Nowakowski TJ, Lee S, Favuzzi E, Kaeser PS, Sjulson L, Batista-Brito R, Takesian AE, Ibrahim LA, Iqbal A, Pelkey KA, McBain CJ, Dimidschstein J, Fishell G, Wang Y. Furlanis E, et al. bioRxiv [Preprint]. 2025 Apr 22:2024.07.17.603924. doi: 10.1101/2024.07.17.603924. bioRxiv. 2025. Update in: Neuron. 2025 May 21;113(10):1525-1547.e15. doi: 10.1016/j.neuron.2025.05.002. PMID: 39091835 Free PMC article. Updated. Preprint.

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An enhancer-AAV toolbox to target and manipulate distinct interneuron subtypes

Affiliations

An enhancer-AAV toolbox to target and manipulate distinct interneuron subtypes

Authors

Affiliations

Abstract

Conflict of interest statement

Update of

MeSH terms

LinkOut - more resources

Full Text Sources

Molecular Biology Databases

Research Materials