C57BL/6J-Pacs1^em2Gath/J

Stock number: 038550
Product name: PACS1^R201W , Pacs1^tm1Gath , Pacs1-LSL-R201W
Research areas: Developmental Biology Research, Mouse/Human Gene Homologs, Neurobiology Research, Research Tools

Description

PACS1^R201W conditional knock-in mice carry an R201W amino acid substitution preceded by a lox-STOP-lox (LSL) cassette in the Pacs1 (phosphofurin acidic cluster sorting protein 1) gene. This strain may be useful for research in neurodevelopmental disorders such as PACS1 syndrome.

Of note, the mutant R203W human PACS1 gene is distributed as C57BL/6J-Gt(ROSA)26Sor^{em2(CAG-PACS1)Gath}/J (Stock No. 038551). In addition, the the human PACS1 control is distributed as C57BL/6J-Gt(ROSA)26Sor^{em2(CAG-PACS1)Gath}/J (Stock No. 038552).

Detailed description

The Pacs1 (phosphofurin acidic cluster sorting protein 1) gene encodes a trafficking protein involved in trans-Golgi network membrane protein localization. A human missense mutation in PACS1, R203W, is associated with developmental delay and intellectual disability (PACS1 syndrome). Functionally, the mutation results in increased dendrite arborization, diminished spine density, and fewer functional synapses.

PACS1^R201W conditional knockin mice carry a CRISPR/cas9 generated amino acid substitution R201W preceded by a lox-STOP-lox (LSL) cassette in the Pacs1 gene. The mutation is orthologous to the human R203W mutation. Mice homozygous for the mutation are believed to be viable and fertile. Upon exposure to Cre recombinase, the LSL is removed and the R201W mutation expressed from the endogenous Pacs1 locus.

When combined with mice carrying cre expressed in neurons (B6.129S2-Emx1^tm1(cre)Krj/J, Stock No. 005628), double mutant progeny exhibit aberrant mEPSCs in cortical neurons and an altered E/I ratio, but had no effect on mIPSCs.

Of note, the mutant R203W human PACS1 gene is distributed as C57BL/6J-Gt(ROSA)26Sor^{em2(CAG-PACS1)Gath}/J (Stock No. 038551). In addition, the the human PACS1 control is distributed as C57BL/6J-Gt(ROSA)26Sor^{em2(CAG-PACS1)Gath}/J (Stock No. 038552).

Development

CRISPR/cas9 endonuclease-mediated genome editing was used to insert a loxP-flanked SA-F-neo-STOP sequence in intron 3, followed by an arginine to tryptophan amino acid substitution at codon 201 (R201W, CGA to TGG and silent ACT to ACC mutation in codon 202) in exon 4 of the Pacs1 locus on chromosome 19. The mutation is orthologous to the human R203W mutation. A single guide RNAs (5'- TAGCTGGACCTGAACCACCAAGG-3’¸ chr19:5,160,141-5,160,163, mouse GRCm38/mm10 assembly), cas9 endonuclease and a PACS1-R201W Megamer were electroporated into C57BL/6J-derived zygotes. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to C57BL/6J mice for germline transmission. The colony was maintained on the C57LB/6J background. Upon arrival, mice were bred to C57BL/6J for at least one generation to establish the colony.

Allele

Symbol: Pacs1^em2Gath
Name: endonuclease-mediated mutation 2, Gary Thomas
MGI accession ID: MGI:7518815
Generation method: Endonuclease-mediated
Attributes: Conditional ready (e.g. floxed); Humanized sequence
Synonyms: Pacs1^{R201W; Pacs1tm1Gath; Pacs1-LSL-R201W
Marker symbol: Pacs1
Marker name: phosphofurin acidic cluster sorting protein 1
Marker synonyms: MGC:37046; MRD17; Pacs-1; SHMS
Chromosome: 19
Strain of origin: C57BL/6J
Molecular note: CRISPR/cas9 endonuclease-mediated genome editing is used to insert a loxP-flanked SA-F-neo-STOP sequence in intron 3, followed by an arginine to tryptophan amino acid substitution at codon 201 (R201W, CGA to TGG and silent ACT to ACC mutation in codon 202) in exon 4 of the gene. The mutation is orthologous to the human R203W mutation.}