To undertake the national key research and development program by using gene-editing canine models

Brief Introduction: In December 2019, Beijing Sinogene Biotechnology co., Ltd. jointly applied for the national key project " key scientific problems in transformative technology" in the "social and emotional biological basic and translational research" projects, which was the national key research plan of Ministry of Science and Technology, with Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Institute of Biophysics, Chinese Academy of Sciences, Kunming Institute of Zoology, Chinese Academy of Sciences and Beijing university. Through the defense, the project was approved successfully.

Project Title: Analyzing the biological basis of social interaction and emotion with the domestic dog mode

Based on the variety of traits, active social interactions, and rich emotions (including cross-species empathy with humans) shown by domestic dogs who were developed over 30,000 years of artificial breeding, Sinogene established the research field of social and emotional and cognitive function in domestic dogs as a model animal, revealed the genetic and neurobiological mechanisms of human social interaction and emotion, and made breakthroughs in the research and development of drugs for cognitive and psychiatric diseases. With the efficient and accurate gene editing and clone technology, Sinogene set up quantity evaluation system for domestic dog’s social, emotional and empathy as well as technology platform for cerebral structure and function analysis to provide ideal model animal for cerebral science research. 

The key science issues to be solved: 

Siongene made full use of the unique nature of domestic dogs' rich social behaviors, including cross-species sociability and empathy with humans, and analyze the key genes and molecular genetic pathways that regulate social and emotional behaviors in domestic dogs using evolutionary genetics and multi-omics methods. Meanwhile, Sinogene constructed social and emotional disorder gene mutation dogs with the CRISPR/Cas9 technology, and the social, emotional and empathic quantitative analysis methods, to reveal how the genetic mutations affect the neurobiological basis of social interaction and emotion through the analysis of the phenotype system, including the molecular and cellular, behavior, social interaction, emotion, electrophysiological and brain imaging analysis. Thus, it provides theoretical guidance for decoding the neural mechanisms of human social interaction and emotion, and for the prevention and treatment of related major mental diseases.

1、build social interaction and emotion quantitative analysis system 

Build social interaction and emotion quantitative analysis system of domestic dogs. Design and establish different experimental paradigms of canine-canine and canine-human social interaction. Analyze eye movement, facial features, tail wagging, physical characteristics of domestic dogs in different scenes, detect domestic dogs’ internal activities and develop video analysis algorithm to quantitatively analyze the social interaction and emotion of mutant dog. Thus, the behavioral analysis and evaluation platform of domestic dogs was established.

2、Multi-omics analysis of key genes and regulatory pathways 

Genes significantly associated with social and emotional interaction were screened by population genetic analysis using high-throughput sequencing platform. Single - cell multi - omics analysis was used to search for related genes at cell level. Through the analysis of transcriptome, lipidome and proteome, the mechanism of the expression of related genes in regulatory pathways was studied.

3、Construction of Gene editing mutant dogs

Canine fetal fibroblasts were cultured in vitro to screen suitable sgRNAs targeting different regions of social and empathy related desired genes, and mutants were prepared using the following two protocols:

4、EEG Analysis

Real-time electrophysiological analysis of brain function was performed in dogs with free mobility. The firing signals and field potential signals of neurons were recorded by multi-channel electrodes to compare the firing activity between mutant and normal animals. Through the coupling analysis of behavior and electrophysiological results, the EEG characteristics and rules of different behaviors and different stimuli are summarized, and the neural mechanism of behavior is decoded.

5、Brain imaging analysis of mutant domestic dogs

Brain structure (MRI) and function (fMRI) were analyzed by magnetic resonance imaging (MRI), and the volume of functional brain regions, nerve fibers linking brain regions, and synchronization of brain regions were compared between normal and mutant dogs, and the effects of target gene mutations on brain structure and function were analyzed. 

In this study, for the first time, canine gene editing technology and cloning technology were used to construct gene mutants related to social interaction and emotion, so as to reveal the biological basis of social interaction and emotion, analyze the pathogenesis of a major mental disorder, namely autism, and provide a key detection system and platform for drug development.