Protocol for inducing severe Scn2a insufficiency in mice by intracerebroventricular antisense oligonucleotide injection

Melody Li; Bernd Kuhn

doi:10.1016/j.xpro.2024.103094

Protocol for inducing severe Scn2a insufficiency in mice by intracerebroventricular antisense oligonucleotide injection

STAR Protoc. 2024 Jun 21;5(2):103094. doi: 10.1016/j.xpro.2024.103094. Epub 2024 May 25.

Authors

Melody Li¹, Bernd Kuhn²

Affiliations

¹ Optical Neuroimaging Unit, Okinawa Institute of Science and Technology Graduate University (OIST), 1919-1 Tancha, Onna-son, Okinawa 904-0495, Japan. Electronic address: [email protected].
² Optical Neuroimaging Unit, Okinawa Institute of Science and Technology Graduate University (OIST), 1919-1 Tancha, Onna-son, Okinawa 904-0495, Japan. Electronic address: [email protected].

Abstract

SCN2A loss-of-function variants cause a range of neurodevelopmental disorders. Here, we present a protocol to induce severe Scn2a insufficiency in mice. We describe steps for intracerebroventricular (ICV) antisense oligonucleotide (ASO) injection that causes a selective downregulation of Scn2a and ASO-mediated mRNA degradation. We then detail procedures for qPCR and western blot protocol to measure Scn2a mRNA and protein. This protocol can be used as a mouse model for behavioral and in vivo two-photon Ca²⁺ imaging.

Keywords: Model Organisms; Molecular Biology; Neuroscience.

MeSH terms

Animals
Disease Models, Animal
Injections, Intraventricular
Mice
NAV1.2 Voltage-Gated Sodium Channel* / genetics
Oligonucleotides, Antisense* / administration & dosage
Oligonucleotides, Antisense* / genetics
Oligonucleotides, Antisense* / pharmacology
RNA, Messenger / genetics
RNA, Messenger / metabolism

Substances

NAV1.2 Voltage-Gated Sodium Channel
Oligonucleotides, Antisense
RNA, Messenger
Scn2a protein, mouse