Overview

These OVE#2495A mice harbor a mutation created by random insertion of the SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223). Using inverse PCR analysis, the transgene integration site was identified in intron 5 of the splicing factor 3a, subunit 1 gene (Sf3a1) on chromosome 11 (specifically at the 5'-4,071,241(+) bp position with the rtTA in sense orientation relative to the endogenous gene). The donating investigator reports the phenotype of homozygous mice as: may be preimplantation lethal.

Donating Investigator

Paul A Overbeek, Baylor College of Medicine

Genetic overview

Genetic Background	Generation

Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}

Allele Type
Transgenic (Transposon concatemer)

View Genetics

Research Applications

Developmental Biology Research

View All Research Applications

Details

Detailed Description

These OVE#2495A mice harbor a mutation created by random insertion of the SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223). Using inverse PCR analysis, the transgene integration site was identified in intron 5 of the splicing factor 3a, subunit 1 gene (Sf3a1) on chromosome 11 (specifically at the 5'-4,071,241(+) bp position with the rtTA in sense orientation relative to the endogenous gene). The donating investigator reports the phenotype of homozygous mice as: may be preimplantation lethal.

Development

A lentiviral transgenic approach was used to generate these mice. The SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223) was designed with a Sleeping Beauty (SB) transposon (containing a slice-acceptor::IRES::rtTA::polyA gene trap), a mouse tyrosinase minigene (Tyro), and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) in the FUGW self-inactivating HIV-based lentiviral vector backbone. The SB transposon and Tyro minigene replaced the ubiquitin-c promoter and EGFP sequences originally found in the FUGW lentiviral vector. The 1200 bp SB transposon used in this transgene has an inverted repeat/direct repeat sequence (IR/DR; the SB transposon recognition site), an adenovirus splice acceptor, a stop sequence (3xSTOP), an internal ribosome entry site (IRES; from human X-chromosome-linked inhibitor of apoptosis (XIAP)), a sequence encoding an optimized form of reverse tetracycline controlled transactivator (rtTA2S; with stop codon), a human growth hormone polyA sequence, and a second IR/DR sequence. The IR/DR sequences are outward-facing (pointed away from the sa-IRES-rtTA-pA). The Tyro minigene is composed of the mouse Tyr enhancer region (623 bp), promoter region (657 bp), and 1566 bp cDNA sequence (including the stop codon); all in sense orientation relative to the FUGW backbone. There is no polyA site between the Tyro minigene and WPRE sequence. The WPRE sequence functions to enhance the mRNA transcript stability. This packaged lentiposon was injected subzonally (under the zona pellucida) into 1-8 cell stage FVB/N mouse embryos. Expression of the tyrosinase minigene results in melanin synthesis, so founder mice (F0) were identified by inspection for pigmentation. All F0 mice exhibited mosaic, light, pigmentation. Most F0 mice had more than one lentiviral integration site. F0 mice were assigned an OVE number and then bred with FVB/N mice. Pigmented offspring (F1) with different coat colors were designated as subline A, B, C, etc., for each family. F1 mice were bred with FVB/N mice. F2 mice with different coat colors were designated as sublines A-1, A-2, etc., or B-1, B-2, etc. Breeding with FVB/N mice was continued until litters with only one or two different coat colors were obtained. Mice with identical coat colors were then inbred and assessed for the presence or absence of viable homozygotes. Hemizygous mice of line OVE2495A (OVE#2495A) exhibit light mottle coat color. Using inverse PCR analysis, the lentiviral integration site was identified in intron 5 of the splicing factor 3a, subunit 1 gene (Sf3a1) on chromosome 11. The 5'-LTR is linked to the (+) strand of DNA at position 4,071,241 bp [NCB137/mm9; 5'-4,071,241(+)]. The rtTA is inserted in the sense orientation relative to the disrupted mouse gene. Transgenic males were used to freeze sperm, and that cryopreserved sperm were sent to The Jackson Laboratory Repository node of the Mutant Mouse Regional Resource Center (MMRRC-JAX). Upon arrival, aliquots of this frozen sperm were used to fertilize oocytes from FVB/NJ inbred females to establish the live colony.

Expression Data

Expressed Gene	Tyr, tyrosinase, mouse, laboratory
Site of Expression

Control Suggestions

001800 FVB/NJ

Additional Information

Considerations for Choosing Controls

Genetics

Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}

Allele Symbol: Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}

Allele Name	transgenic transposon lentiviral insertion 2495A, Paul A Overbeek
Allele Type	Transgenic (Transposon concatemer)
Allele Synonym(s)	OVE#2495A; OVE2495A
Gene Symbol and Name	Sf3a1, splicing factor 3a, subunit 1
Gene Synonym(s)
Promoter	Tyr, tyrosinase, mouse, laboratory
Expressed Gene	Tyr, tyrosinase, mouse, laboratory
Strain of Origin	FVB/N
Chromosome	11
Molecular Note	A lentiviral transgenic approach was used to generate these mice. The SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223) was designed with a Sleeping Beauty (SB) transposon (containing a slice-acceptor::IRES::rtTA::polyA gene trap), a mouse tyrosinase minigene (Tyro), and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) in the FUGW self-inactivating HIV-based lentiviral vector backbone. The SB transposon and Tyro minigene replaced the ubiquitin-c promoter and EGFP sequences originally found in the FUGW lentiviral vector. The 1200 bp SB transposon used in this transgene has an inverted repeat/direct repeat sequence (IR/DR; the SB transposon recognition site), an adenovirus splice acceptor, a stop sequence (3xSTOP), an internal ribosome entry site (IRES; from human X-chromosome-linked inhibitor of apoptosis (XIAP)), a sequence encoding a-n optimized form of reverse tetracycline controlled transactivator (rtTA2S; with stop codon), a human growth hormone polyA sequence, and a second IR/DR sequence. The IR/DR sequences are outward-facing (pointed away from the sa-IRES-rtTA-pA). The Tyro minigene is composed of the mouse Tyr enhancer region (623 bp), promoter region (657 bp), and 1566 bp cDNA sequence (including the stop codon); all in sense orientation relative to the FUGW backbone. There is no polyA site between the Tyro minigene and WPRE sequence. The WPRE sequence functions to enhance the mRNA transcript stability. In line OVE#2495A, a single copy of the lentiposon inserted into intron 5 [NCBI37/mm9; 5'-4,071,241(+)] in the sense orientation.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Developmental Biology Research
- Embryonic Lethality (Homozygous)

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}/Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}
FVB/N-Sf3a1

mortality/aging

embryonic lethality before implantation
- may be preimplantation lethal

References

Additional - Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve} related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Separated PCR:Sf3a1-3'
- Separated MCA:Sf3a1 3'
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred together, bred with wildtype siblings, or bred with FVB/N inbred mice.
- Additional Breeding and Husbandry Support
Mating System
- Wild-type x Heterozygote
- Heterozygote x Wild-type
Citation
When using the FVB/N-Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #36217 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Terms Of Use

General Terms and Conditions

See MMRRC for Additional Conditions of Distribution

Questions about Terms of Use

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

JAX® Mice, Products & Services Conditions of Use

"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.

No Warranty

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.

Credit for PRODUCTS or SERVICES

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.

Animal Care and Use for SERVICES

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.

No Liability

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.

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Sf3a1TgTn(sb-rtTA,Tyr)2495AOve

Allele Symbol: Sf3a1TgTn(sb-rtTA,Tyr)2495AOve

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Sf3a1TgTn(sb-rtTA,Tyr)2495AOve/Sf3a1TgTn(sb-rtTA,Tyr)2495AOveFVB/N-Sf3a1

mortality/aging

References

Additional - Sf3a1TgTn(sb-rtTA,Tyr)2495AOve related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}

Allele Symbol: Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}

Genotype: Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}/Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve}
FVB/N-Sf3a1

Additional - Sf3a1^{TgTn(sb-rtTA,Tyr)2495AOve} related