Creating a STAT2 Knock-Out Rat With CRISPR/Cas9 Using i-GONAD

Author

Isabella DeJesus, Utah State University

Date of Award

5-2025

Degree Type

Thesis

Degree Name

Departmental Honors

Department

Animal, Dairy, and Veterinary Sciences

Abstract

Through the use of genetically modified animals, a more holistic understanding of physiologic responses to various diseases and pathologies can be obtained^7,19. The STAT2 gene encodes a protein called Signal Transducer and Activator of Transcription 2 and is an excellent target for genetic manipulation due to its essential role in type I and III interferon signaling pathways¹¹. A member of the STAT family of transcription factors, this protein is directly involved in mediating cellular responses to various cytokines and growth factors, particularly those related to the innate immune response¹¹. While the STAT2 signaling pathway is frequently studied, no verifiable creation of a rat STAT2 knock-out (KO) has been observed in the literature. By establishing an effective procedure for creating STAT2 KO rats, these organisms can be used for refined disease testing of serious human ailments such as Lupus, Polio, Herpes, RSV, and even cancer^{1,3,9,11,12,20}. In this paper, we, the Wang Lab in the department of Animal, Dairy, and Veterinary Sciences at Utah State University illustrate a protocol for creation of the first ever STAT2 KO Dawley Rat using CRISPR/Cas9 and i-GONAD techniques. Through highly specific sgRNA design, detailed tracking of estrus cycles for optimal timing of i-GONAD, and numerous genotyping procedures such as PCR-RFLP and TA cloning, successful STAT2 homozygous KO rats were generated. The development of these gene edited animals will have a significant positive impact on the ability to study immune related diseases as well as increase the translational ability of application study results for human consideration.

Recommended Citation

DeJesus, Isabella, "Creating a STAT2 Knock-Out Rat With CRISPR/Cas9 Using i-GONAD" (2025). Undergraduate Honors Capstone Projects. 1027.
https://digitalcommons.usu.edu/honors/1027