Reuben Ryan Cano

Reuben Ryan Cano, a senior studying pre-med biology at the University of Utah, validated two mouse models for the functional analysis of a schizophrenic risk gene CACNA1I.

Schizophrenia is a debilitating mental disorder that affects about 1% of the world population. The etiology of schizophrenia is largely unknown, making it difficult to diagnose and to develop targeted therapies. At the Broad I was surrounded by world-leading scientists who were as brilliant as they were humble and supportive. Not only was I able to learn from them to develop as a scientist, but I also grew as a person. I learned to cast my doubts aside and realized that I, too, am capable of doing great science that might one day have an impact on eradicating diseases and ameliorating suffering. I also learned how I could combine my goal of alleviating health disparities with my passion for research as I pursue a career as a physician scientist.Despite the challenges, recent genome-wide association studies have identified possible genetic drivers of this disease. One of these genes, CACNA1I, encodes a T-type calcium channel in neurons critical for sleep spindle generation. Sleep spindle deficits are pronounced in schizophrenia patients, but due to lack of specific antibodies, little is known about the localization of this calcium channel within the cells and brain circuitries it is involved. To circumnavigate this problem, we developed two mouse models with knocked-in C-terminal tags, P2A-Cre and Clover (a GFP variant). Through PCR and Sanger sequencing, we confirmed both reporters were knocked in correctly on the endogenous CACNA1I gene. We further validated the Cav3.3-Clover model by several biochemical assays. Using Western blotting, we confirmed the fusion protein remains intact after expression, which enabled us to use GFP as a valid indicator of CaV3.3’s location. We were then able to show that Cav3.3 is localized in dendrites, but not at excitatory synapses. These animal models will provide valuable insights into the localization of this calcium channel and its function. Such insights, in turn, could provide valuable understanding of how dysfunction of this channel might lead to the development of schizophrenia, eventually leading to identifying potential biomarkers of the disease for diagnosis and patient stratification, as well as developing better therapeutic approaches for schizophrenic patients.

Project: Knock-in reporter models to annotate the function of CACNA1I

Mentor: Lingling Yang, Stanley Center for Psychiatric Research