services for the Generation of THRB-Cre-ERT2 Knock-In Mouse

PRE-SOLICITATION NON-COMPETITIVE (NOTICE OF INTENT) SYNOPSIS INTRODUCTION THIS IS A PRE-SOLICITATION NON-COMPETITIVE (NOTICE OF INTENT) SYNOPSIS TO AWARD A CONTRACT OR PURCHASE ORDER WITHOUT PROVIDING FOR FULL OR OPEN COMPETITION (INCLUDING BRAND-NAME). The National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), Office of Acquisitions (OA) on behalf of the on behalf of the National Institute of Digestive, Diabetes & Kidney Diseases intends to award a purchase order without providing for full and open competition (Including brand-name) to OZGENE PTY LTD for services for the Generation of THRB-Cre-ERT2 Knock-In Mouse. NORTH AMERICAN INDUSTRY CLASSIFICATION SYSTEM (NAICS) CODE The intended procurement is classified under NAICS code 541380 with a Size Standard of $16.50. REGULATORY AUTHORITY The resultant contract will include all applicable provisions and clauses in effect through the Federal Acquisition Circular 2022-04 Effective January 30, 2022. This acquisition is conducted under the procedures as prescribed in FAR subpart 13'Simplified Acquisition Procedures at an amount not exceeding the simplified acquisition threshold ($250,000). STATUTORY AUTHORITY This acquisition is conducted under the authority of 41 U.S.C. 253(c) under provisions of the statutory authority of FAR Subpart 6.302- FAR 6.302-1'Only one responsible source and no other supplies or services will satisfy agency requirements 41 U.S.C. 253(c)(1). PERIOD OF PERFORMANCE Within 12 Months of Date of Award Place of Performance National Institutes of Health National Institute of Digestive, Diabetes & Kidney Diseases 50 South Drive Bethesda, MD 20892 DESCRIPTION OF REQUIREMENT Thyroid hormone is important for tissue development and for maintaining general health in adulthood. There are two thyroid hormone receptors alpha and beta. The beta receptor is encoded by the THRB gene and has two spliced variants, THRB1 and THRB2. They have their unique exon1 but share common downstream exons. These receptors express differently in various tissues. Our previous mouse models containing the mutation of these receptors have various sensory and developmental defects, demonstrating the importance of these receptors. To understand more precisely the tissues and cell types in which THRB1 functions, it is important to be able to trace the expression pattern of the THRB1 protein. As there is no antibody available that reliably detects THRB1 protein in natural tissues, we propose to create a mouse model which carries a THRB1-Cre-ERT2 cassette. This model will allow sensitive detection of THRB1 protein in mouse tissues. The Flpe delete mice will be crossed onto the new mouse model to remove the neo selection cassette in the mutated genome of the model mouse. The goal of the work is to generate a mouse strain that carries a specific, heritable alteration in the THRB1 gene. The task involves the application of gene targeting technology of Cre-ERT2 recombinase to introduce the mutation into the desired gene. This is performed in mouse embryonic stem cells which will be used to create founder mice carrying the targeted mutation in the germ-line. _________________________ PURPOSE AND OBJECTIVES The objective of this request is to procure services to generate a mouse strain that carries a specific, heritable alteration in the THRB1 gene. The task involves the application of gene targeting technology of Cre-ERT2 recombinase to introduce the mutation into the desired gene. This is performed in mouse embryonic stem cells which will be used to create founder mice carrying the targeted mutation in the germ-line. SALIENT / REQUIRED FEATURES AND SPECIFICATIONS The work includes the following steps: 1. Design and construction of the required targeting vector. All steps should be verified at the highest standard of proof for having been correctly performed. 1A. The vector should carry adequate lengths of homologous sequence on either side of the alteration to give a high probability of correct integration at the targeting site in the genome. Due attention should be paid to the mouse strain of origin of the homologous sequences to optimize the probability of achieving a specific targeted integration as opposed to non-homologous random integration in the genome. 1B. The vector should incorporate standard elements including a neomycin-resistance marker and flanking sites (FRT) that allow for the later excision of the marker after successful targeting of the gene has been achieved. 1C. The vector will contain the Cre-ERT2 cassette inserted downstream of ATG start-site in the unique exon 1 of THRB1 gene. 2. Selection of recombinant ES cell clones carrying the correctly targeted insertion at the gene of interest. The linearized targeting vector should be introduced into the ES cells and colonies carrying the integrated vector should be selected based on criteria including neomycin-resistance and direct demonstration of correct homologous recombination by Southern blot analysis of genomic DNA. 3. Generation of mice carrying the mutation. Correctly targeted ES cell clones with correct karyotypes should be used to generate founder mice by injection of the cells into blastocysts of a host mouse strain. Transmission of the mutation by the founder mice should be demonstrated following backcrossing and direct demonstration by Southern blot analysis of genomic DNA. The provider should have demonstrated experience with appropriate technology to remove the neomycin marker while retaining the desired alteration at the targeted gene. 4. The founder mice will be crossed onto Flpe-deleter mice to remove the neomycin-cassette in their genome. Requirements 1. Expertise in the field of gene targeting technology and a proven record of accomplishment. The provider should have been successful for a significant number of years in this field. Evidence for proven success includes the publication of a significant number of successful projects in reputable, peer-reviewed scientific journals. 2. Proven experience in performing similar projects in embryonic stem cell lines of different mouse strain origin. This includes the option of using C57BL/6- or 129 substrain-derived ES cells, each with high probabilities of success, supported by evidence from publications in peer-reviewed scientific journals. 3. Ability to perform the tasks within a competitive timeframe for an important scientific project. Expertise in techniques that enhance the efficiency of the service, such as optimized techniques using ES cells that produce germline transmission of the mutation with least number of animals used. 4. Provision of services for a competitive price. TASKS AREAS Independently, and not as an agent of the Government, the contractor shall perform all of the detailed Task Areas listed below: Task Area 1 - Vendor will provide 2-4 pairs of breeding mice carrying the TRB1Cre mutation. Quote with estimated shipping costs attached to order. The delivery timeline for this order is 30-40 weeks. Task Area 2 - When the mice are ready to ship, the vendor will coordinate with our institute's animal import officer. Vendor will arrange shipment including the export permit and contacting a specialized courier. They will use approved animal containers and pack the animals with specialized food for transportation. We will pay for the extra shipping fee at that time. Task Area 3 - Vendor will keep a back-up colony. In the case of any death during transit, the vendor will ship more animals to us. Task Area 4 - We will perform PCR to genotype the mice to ensure the animals carry the targeted mutation. CONTRACTING WITHOUT PROVIDING FOR FULL OR OPEN COMPETITION (INCLUDING BRAND-NAME) DETERMINATION The determination by the Government to award a contract without providing for full and open competition is based upon the market research conducted as prescribed in FAR Part 10'Market Research. Only one source is available: Per FAR 13.106-1(b)(1) the Contracting Officer has determined OZGENE PTY LTD to be the only reasonable available source to provide services for the Generation of THRB-Cre-ERT2 Knock-In Mouse. This acquisition was pursued on a sole source basis centered on the following. There is no commercial antibody available that reliably detects THRB1 protein in natural tissues. Currently, in-situ hybridization is the only technique to investigate the expression of THRB gene in fixed tissues. The selected offsprings will then be crossed onto commercially available reporter Ai6 mice from the Jackson Lab. The hybrid mice will allow sensitive detection of THRB1 protein because they will contain the green fluorescent THRB-GFP protein. Thus, OZGENE PTY LTD is the only vendor that can fulfill this requirement. Ozgene has the ability to design and construct efficient targeting vectors specifically to engineer the - Cre-ERT2 cassette at exon1 of THRB1 gene but not THRB2 gene so that the new mouse model carries mutation only in THRB1 but not the other isoform THRB2 (THRB1 and THRB2 are spliced variants, different only in their unique exon 1 but share downstream exons). In addition, the NIDDK Investigator has previously used the same technology as provided by Ozgene, which provides an advantage of consistency between different projects in the Investigator's lab. This allows certain analyses to be compared meaningfully between different projects. The investigator has used a knock-in model mouse (POTS # 19-003228) created by Ozgene, which resulted in a recent publication in the journal Science Advances 10 Jun 2020: Vol. 6, no. 24, eaba2634. CLOSING STATEMENT This synopsis is not a request for competitive proposals. However, interested parties may identify their interest and capability to respond to this notice. Responses to this notice shall contain sufficient information to establish the interested parties bona-fide capabilities for fulfilling the requirement and include: descriptive literature, delivery timeframe, warranties and/or other information that demonstrates that the offer meets all the foregoing requirements, the prompt payment discount terms, the F.O.B. Point (Destination or Origin), the Dun & Bradstreet Number (DUNS), the Taxpayer Identification Number (TIN), and the certification of business size. All offerors must have an active registration in the System for Award Management (SAM) www.sam.gov. A determination by the Government not to compete this proposed contract based upon responses to this notice is solely within the discretion of the Government. The information received will normally be considered solely for the purposes of determining whether to proceed on a non-competitive basis or to conduct a competitive procurement. All responses to this notice shall be submitted electronically by 9:00 am Eastern Standard Time, on Thursday, February 10, 2022 to the Contract Specialist, Amber Harris, at contact@example.gov . Assessment of Capability Lowest Price Technically Acceptable