These Shank3(+/ΔC) mice may be useful for studying behavioral and synaptic abnormalities associated with autism spectrum disorder.
Paul Worley, Johns Hopkins University School of Medic
These Shank3(+/ΔC) mutant mice lack exon 21 of the SH3/ankyrin domain gene 3 (Shank3) gene. The removal of exon 21 causes a frameshift which results in the production of SHANK3 protein lacking the entire C-terminal region, including the Homer-binding site in the sterile alpha motif (SAM) domain. SHANK3 is a synaptic scaffolding protein expressed in the postsynaptic density (PSD) of excitatory synapses. SHANK3 mutations have been identified in cases of intellectual disability such as autism spectrum disorder (ASD), Phelan-McDermid syndrome, and schizophrenia. Truncated SHANK3 protein associates with wildtype protein resulting in a 75% reduction in wildtype SHANK3 in cortical cultures, and 90% reduction in synaptic cultures. These Shank3(+/ΔC) mice exhibit behavioral deficits suggestive of autism, including aberrant social interactions leading to aggressive behaviors. These mice also have reduced NMDA receptor function in hippocampal neurons resulting in the reductions in long term potential and depression. Shank3(+/ΔC) mice display normal learning and memory, and show no alterations in postsynaptic density proteins, spine morphology, or synapse number. Mice that are heterozygous for this allele are viable and fertile. The donating investigator has reported some fertility issues when maintaining a homozygous colony.
A targeting vector was designed to insert a loxP site upstream of exon 21 followed by a frt-flanked neomycin resistance (neo) cassette, and a second loxP site downstream of exon 21 of the SH3/ankyrin domain gene 3 (Shank3) gene. The construct was electroporated into 129S6/SvEvTac embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and resulting chimeric mice were bred to C57BL/6J mice. The resulting offspring, were bred with mice expressing actin-cre to delete the neo cassette and exon 21, and progeny were crossed to remove the Cre-expressing transgene. The donating investigator reported that the resulting Shank3(ΔC) mice were bred to C57BL/6J mice for at least 5 generations (see SNP note below). Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. 18 of the 27 markers throughout the genome were found to be segregating for 129 and B6. Also, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.
|Allele Name||targeted mutation 1.1, Paul Worley|
|Allele Type||Targeted (Hypomorph)|
|Allele Synonym(s)||Shank3(deltaC); Shank3tm1.1Bxia|
|Gene Symbol and Name||Shank3, SH3 and multiple ankyrin repeat domains 3|
|Strain of Origin||129S6/SvEvTac|
|General Note||The data reported for this allele were originally described in: |
Bangash MA, Park JM, Melnikova T, Wang D, Jeon SK, Lee D, Syeda S, Kim J, Kouser M, Schwartz J, Cui Y, Zhao X, Speed HE, Kee SE, Tu JC, Hu JH, Petralia RS, Linden DJ, Powell CM, Savonenko A, Xiao B, Worley PF. Enhanced polyubiquitination of Shank3 and NMDA receptor in a mouse model of autism. Cell. 2011 May 27;145(5):758-72. PMID:21565394.
This paper was retracted; however, the prinicpal investigator has confirmed the validity of the data related to the molecular gene targeting and the phenotypic description of the mutant mice carrying this allele. Please see the retraction notes for further details about the data in the original paper.
|Molecular Note||A loxP site was inserted upstream of exon 21. An FRT-flanked neo cassette with a 3' loxP site was inserted downstream of exon 21. Cre-mediated recombination removed exon 21 and the neo cassette. No wild-type protein is detected in homozygous mice but a lower molecular weight mutant protein is detected by western blot analysis.|
|Mutations Made By|| |
Bo Xiao, Johns Hopkins University School of Medic
When maintaining a live colony, heterozygous males may be bred to wildtype females from the colony or to C57BL/6J female mice (Stock No. 000664). The donating investigator maintains this non-reciprocal cross to exclude effects of possible maternal autism spectrum disorder (ASD)-like phenotype on rearing. Some fertility issues have been reported when maintaining a homozygous colony.
When using the Shank3ΔC (Shank3Δ ex21) mouse strain in a publication, please cite the originating article(s) and include JAX stock #018398 in your Materials and Methods section.
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCT(S)" means biological materials supplied by JACKSON, and their derivatives. "SERVICES" means projects conducted by JACKSON for other parties that may include but are not limited to the use of MICE or PRODUCTS. "RECIPIENT" means each recipient of MICE, PRODUCTS, or SERVICES provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE, PRODUCTS or SERVICES from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON’s prior written authorization.
MICE, PRODUCTS AND SERVICES ARE PROVIDED "AS IS". JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of, PRODUCTS or SERVICES, JACKSON will, at its option, provide credit or replacement for the PRODUCT received or the SERVICES provided; JACKSON makes no other representations and this shall be the exclusive remedy of the purchaser. Please note specific policy for live mice.
Consistent with the requirement for a written understanding regarding animal care and use, the JACKSON Animal Care and Use Committee will review the animal care and use protocol(s) associated with any SERVICES to be performed at JACKSON, and JACKSON shall have ultimate responsibility and authority for the care of animals while on site or in JACKSON custody.
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS, or SERVICES, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS, or SERVICES from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE, PRODUCTS or SERVICES are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or SERVICES. In addition, special terms and conditions of sale of certain MICE, PRODUCTS, or SERVICES may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and SERVICES by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or SERVICES shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or SERVICES by JACKSON.