Arid1b Flox (Arid1bFl; floxed exon 5) mice have a CRISPR/cas9-generated, Cre-conditional knock-out allele. These mice may be useful in studying the SWI/SNF chromatin-remodeling complex, autism spectrum disorder, intellectual disability, corpus callosum agenesis and Coffin-Siris syndrome.
Hao Zhu, University of Texas Southwestern Medical Center [UTSW]
ARID1B is a SWI/SNF chromatin-remodeling subunit. ARID1B haploinsufficiency in humans is associated with autism spectrum disorder, intellectual disability, corpus callosum agenesis and short stature. In addition, ARID1B mutation is the most common cause of Coffin-Siris syndrome.
The Arid1b Flox allele (Arid1bFl) has loxP sites flanking exon 5. Mice homozygous for this floxed allele are viable and fertile, with no reported gross phenotypic or behavioral abnormalities.
Upon exposure to Cre recombinase, the floxed sequences are deleted - resulting in a null allele (Arid1bΔ).
For example, when Arid1b Flox are bred to Nestin-Cre transgenic mice (e.g., Stock No. 003771), the resulting brain-specific ARID1B haploinsufficiency (Nestin-Cre; Arid1bFl/+) leads to growth impairment and reduced plasma insulin-like growth factor (IGF1) levels with inappropriate lack of growth hormone (GH) increase - characteristics of ARID1B mutation in humans. [Celen et al. 2017 Elife 6:e25730]
Furthermore, breeding Arid1b Flox to mice with liver-specific or skeletal muscle-specific Cre-expression (Stock Nos. 003574 or 006475, respectively) generates mice with tissue-specific ARID1B haploinsufficiency. In the resulting offspring, neither Albumin-Cre; Arid1bFl/+ nor Ckmm-Cre; Arid1bFl/+ showed growth or morphological defects. [Celen et al. 2017 Elife 6:e25730]
In addition, breeding Arid1b Flox to germline Cre-expressing mice (e.g., Sox2-Cre; Stock No. 008454) should generate mice with the ARID1B global knock-out allele. Mice homozygous or heterozygous for the global knock-out allele can be expected to have the same phenotype as other ARID1B whole-body knock-out alleles. That is, homozygotes are perinatal lethal, while heterozygotes are viable and fertile with social behavior impairment, altered vocalization, anxiety-like behavior (increased self-grooming), neuroanatomical abnormalities and growth impairment. Approximately 7% of mice heterozygous for a whole-body knock-out of the gene also have hydrocephaly.
The Arid1b Flox allele (Arid1bFl; floxed exon 5) was created in the laboratory of Dr. Hao Zhu (University of Texas Southwestern Medical Center) using CRISPR/cas9 endonuclease-mediated genome editing. Guide RNAs were designed to target sequences before and after exon 5 of the AT rich interactive domain 1B [SWI-like] locus (Arid1b) on chromosome 17. The guide RNAs, oligo donors containing loxP sequences and S. pyogenes CRISPR/cas9 mRNA were injected into single-celled C57BL/6J zygotes. Founder mice with the Arid1b Flox allele (loxP::exon5::loxP) were identified by PCR and sequencing of the Arid1b exon5 region. Founders were bred to C57BL/6J mice for germline transmission. The Arid1b Flox colony was backcrossed to C57BL/6J for approximately five generations, and then bred together as homozygotes. To date (May 2018), these mice have not been characterized for off-target mutations. In 2018, males with black coat color were sent to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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. One of the 27 markers throughout the genome were segregating with 129. Also, all 4 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating with an unknown source. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.
|Allele Name||endonucelase mediated mutation 2, Hao Zhu|
|Allele Type||Endonuclease-mediated (Conditional ready (e.g. floxed))|
|Gene Symbol and Name||Arid1b, AT rich interactive domain 1B (SWI-like)|
|Strain of Origin||C57BL/6J|
|Molecular Note||LoxP sites were inserted surrounding exon 5.|
Mice homozygous for the Arid1b Flox allele are viable and fertile, with no reported gross phenotypic or behavioral abnormalities. When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). Alternatively, homozygous mice may be bred together.
When using the Arid1b Flox (Arid1bFl; floxed exon 5) mouse strain in a publication, please cite the originating article(s) and include JAX stock #032061 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.