CRISPR Generated Gene-Knockout Mutants for Studying Nicotine-Seeking and Avoidance Behavior in Zebrafish

CRISPR Generated Gene-Knockout Mutants for Studying Nicotine-Seeking and Avoidance Behavior in Zebrafish

Document Type

Poster

Publication Date

10-1-2025

Abstract

The use of tobacco products remains one of the leading causes of preventable diseases worldwide and in the United States. Although the use of tobacco products in the United States declined about 28 million adults in the United States still smoke. Both environmental and genetic factors contribute to nicotine use behavior. To identify targets for potential pharmacotherapeutics that could lower nicotine use behavior, we used a nicotine seeking and avoidance behavioral assay to test larval zebrafish with nonfunctional acetylcholine receptor subunit genes (knockout genes) that we generated in our lab using CRISPR. Genotyping of individual zebrafish was used to identify mutants with one (heterozygous) or two (homozygous) mutant alleles. Off-spring from homozygous mutants was used for behavioral tests. The knockout of different acetylcholine receptor genes resulted in differential shifts of nicotine seeking behavior of larval zebrafish related to cumulative time spent in the nicotine compartment and to tested nicotine concentrations. The gene knockouts also impacted the percentages of nicotine seeking and avoiding larval zebrafish slightly. Overall, the results indicate that acetylcholine receptor subunits play a role in nicotine-seeking and avoidance behavior in larval zebrafish.

Department

Department of Biology, DePauw University IN

Project Mentor

Henning Schneider

Funding and Acknowledgements

Science Research Fellows program

CRISPR Generated Gene-Knockout Mutants for Studying Nicotine-Seeking and Avoidance Behavior in Zebrafish

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