Center of Excellence in Addiction Studies
Genetic Targeting of Neural Circuits Core
Traditional neuropharmacology does not allow identification of connections between activity of defined neurons or neural circuits and behaviors leading to addiction. The emergence of novel genetic techniques enables expression of engineered cell surface ion channels and receptors that can be manipulated with high temporal and spatial resolution by light or by exogenous compounds. CRISPR/Cas9 approaches allow cell specific editing of native proteins, including neurotransmitter receptors. In combination with Cre-Lox expressing mouse lines and viral targeting strategies, these novel techniques are invaluable in interrogating the functional roles of brain addiction circuits. The Genetic Targeting Core provides services to target and manipulate addiction-related mechanisms in a cell and circuit specific manner using state-of-the-art genetic techniques that include:
- Optogenetics is a field of neuroscience that involves the use of light to control the activity of genetically modified neurons. It involves the expression of light-sensitive proteins called opsins in specific neurons, which allows researchers to activate or inhibit these neurons with precise temporal and spatial control using light.
- One current use of optogenetics is in the study of neural circuits and their role in behavior. By activating or inhibiting specific neural populations, researchers can gain insight into the function of these circuits and how they contribute to the expression of various behaviors.
- Optogenetics has also been used in the treatment of neurological disorders. For example, optogenetic therapy has been shown to be effective in reducing the symptoms of Parkinson's disease and in restoring vision in mice with retinal degeneration.
- There is also potential for optogenetics to be used in the development of brain-machine interfaces, which could allow individuals with paralysis to control prosthetics or other assistive devices with their thoughts.
- Overall, optogenetics represents a powerful tool for understanding the brain and has the potential to revolutionize the treatment of neurological disorders.
- Chemogenetics is a field of neuroscience that involves the use of small molecules to control the activity of genetically modified neurons. It involves the expression of receptors that are activated by specific small molecules in specific neurons, which allows researchers to activate or inhibit these neurons with precise temporal and spatial control using these molecules.
- One current use of chemogenetics in the field of addiction is in the study of neural circuits underlying drug seeking and drug taking behaviors. By activating or inhibiting specific neural populations, researchers can gain insight into the function of these circuits and how they contribute to the expression of these behaviors. This knowledge can be used to develop targeted therapies for the treatment of addiction.
- Chemogenetics has also been used to study the neural circuits underlying the rewarding effects of drugs of abuse, such as opiates. By inhibiting specific neural populations, researchers have been able to reduce the rewarding effects of opiates in animal models. This suggests that chemogenetic therapies may be effective in reducing the likelihood of relapse in individuals with opiate addiction.
- There is also potential for chemogenetics to be used in the development of new therapies for the treatment of opiate addiction. For example, chemogenetic therapies that selectively target specific neural circuits may be able to reduce the negative consequences of opiate withdrawal, such as pain and discomfort, and improve the likelihood of successful recovery.
- Overall, chemogenetics represents a powerful tool for understanding the neural basis of addiction and has the potential to revolutionize the treatment of substance use disorders.
- CRISPR/Cas9 is a powerful tool for gene editing that allows researchers to make precise changes to the genome of living cells. It works by using a small RNA molecule to guide an enzyme called Cas9 to a specific location in the genome, where it can cut the DNA at that location. This cut can then be repaired by the cell's own repair machinery, leading to the introduction of a desired change in the genome.
- One potential use of CRISPR/Cas9 in the field of addiction is in the study of the genetic basis of substance use disorders. By making precise changes to the genome of animal models, researchers can study the effects of these changes on behaviors related to drug seeking and drug taking. This can provide insight into the genetic factors that contribute to addiction and may lead to the development of targeted therapies for the treatment of addiction.
- CRISPR/Cas9 has also been used to study the genetic basis of the rewarding effects of drugs of abuse, such as opiates. By making precise changes to the genome of animal models, researchers have been able to modify the rewarding effects of opiates. This suggests that CRISPR/Cas9-based therapies may be effective in reducing the likelihood of relapse in individuals with opiate addiction.
- There is also potential for CRISPR/Cas9 to be used in the development of new therapies for the treatment of opiate addiction. For example, CRISPR/Cas9-based therapies that target specific genes may be able to reduce the negative consequences of opiate withdrawal, such as pain and discomfort, and improve the likelihood of successful recovery.
- Overall, CRISPR/Cas9 represents a powerful tool for understanding the genetic basis of addiction and has the potential to revolutionize the treatment of substance use disorders.
The genes for optogenetic and chemogenetic manipulation as well as CRISPR/Cas9 editing can be delivered into specific cell types and neural circuits using a variety of gene delivery and expression vectors in combination with different genetic mouse lines.
Optogenetic strategies use microbial-based light activated ion channels (opsins) that allow fast neuronal activation (channelrhodopsins) or inhibition (halorhodopsins, archaerhodopsins). Chemogenetic approaches use engineered ion channels (PSAMs) or G protein-coupled receptors (DREADDs) that can be pharmacologically activated. CRISPR/Cas9 gene editing allows direct in vivo manipulation of the genome in rodents. The genes for these proteins can be delivered into specific cell types and neural circuits using a variety of gene delivery and expression vectors in combination with different genetic mouse lines.
The Porreca/Navratilova group investigates the peripheral and central mechanisms of acute and chronic pain and pain-related comorbidities including sleep disruption and cognitive impairments. Additionally, we study opioid circuits promoting pain relief as well as opioid addiction.
What do we offer?
- We provide assistance with the selection and design of gene targeting strategies.
- We will perform pilot studies to develop and optimize specific gene delivery protocols, using all appropriate controls to ensure the lack of pathogenicity and efficiency and specificity of the approach.
- We will develop suitable methods including fluorescent microscopy, western blot analysis, and ELISA to validate the selected approach and to verify and quantify gene targeting efficacy.
Edita Navratilova, PhD
Assistant Professor, Pharmacology
Frank Porreca, PhD
Associate Department Head, Pharmacology
Co-Director, Center for Substance Abuse Research