Snca knockout mice

Adult C57BL/6J mice, Snca‐knockout mice, and human A53T variant α‐syn transgenic line M83 were from the Jackson Laboratory (stock number: 000664, 003692 and 004479, respectively). M83 mice were backcrossed with C57BL/6J mice for at least 10 generations to obtain TgA53T mice with C57BL6/J congenic strain. Then the heterozygous male and female TgA53T mice were bred to obtain homozygous offspring and wild‐type littermates. The genotypes were identified using RT‐PCR. Animal maintenance and experiments were performed in accordance with the Declaration of Helsinki and guidelines of Renmin Hospital of Wuhan University. The protocol was reviewed and approved by the Animal Care and Use Committee of Renmin Hospital of Wuhan University (20210103).

All animal experiments were approved by the Van Andel Institute Institutional Animal Care and Use Committee (IACUC) and conducted in strict accordance with the NIH Guild for the Care and Use of Laboratory Animals. Rodents were provided with food and water ad libitum, exposed to a 12 h light/dark cycle and maintained in a pathogen-free barrier facility. Female adult Sprague-Dawley rats (weighing ~ 180–200 g) were obtained from Charles River Laboratories and used for the stereotaxic delivery of AAV vectors. VPS35^FLOX/WT mice carrying a floxed “WT mini-gene” insertion that disrupts VPS35 expression (Vps35^tm1.2Mjff, stock no. 021807) were obtained from The Jackson Laboratory and described previously [30 ]. SNCA knockout mice (with deletion of exons 1–2; Snca^tm1Rosl, stock no. 003692) were obtained from The Jackson Laboratory, and human A53T-α-Syn transgenic mice (line G2-3, driven by a mouse prion protein promoter; stock no. 006823) have been described [34 (link)]. Mice were identified by genomic PCR using established genotyping protocols. Double mutant mice (A53T-α-Syn^Tg/+/VPS35^FLOX/WT) and appropriate littermate controls were generated by a single round of crossbreeding between hemizygous human A53T-α-Syn and heterozygous VPS35^FLOX/WT mice.