A--MODEL CREATION STRATEGIES FOR Sptbn1-CRE"er (GENE ID: 20742 Tgfbr2-CRE/ER (TBR-II) Gene ID: 21813 MOUSE GENETICS DEVELOPMENT SERVICE VETERAN AFFAIRS MEDICAL CENTER, WASHINGTON DC

Page 1 of 4 SOURCES SOUGHT NOTICE: THIS IS A SOURCE SOUGHT NOTICE/REQUEST FOR INFORMATION (RFI) ONLY. THIS IS NOT A REQUEST FOR PROPOSAL AND/OR QUOTE. The Washington DC VA Medical Center Research Department is performing market research to determine if there are sufficient number of qualified (1) Service Disabled Veteran Owned Small Business (2) Veteran Owned Small Business (3) Small Business set asides to provide the following: Project: Model Creation Strategies or Sptbn1-CREER (Gene ID: 20742) and Tgfbr2-CREER (TBR-II) Gene ID: 21813 Services required are the delivery of at least 2 confirmed F1 mice as well as supplies for genotyping and other information needed about the generation of a Tgfbr2-CREER and a Sptbn1-CREER knock-in/knockout mouse strain (the founder mice). Located at the Department of Veterans Affairs Medical Center, Washington DC (DC VAMC). The contractor must deliver reagents enabling generation of a Sptbn1-CREER knockin/out strain and Tgfbr2-CRE knockin/out strain at the end of Milestone 2, which has an estimated time of 12-16 weeks. Additionally, after Milestone 4, the shipment of at least two confirmed F1 mice must be executed in a timely manner following the 8-10 weeks estimated time needed to conduct this final Milestone. DC-VAMC is seeking sources from contractors in the Metropolitan Washington DC, Maryland and Virginia area. All interested parties include Large and small businesses as well as Woman-owned Small Business (WOSB) are welcome to respond in the event no Service Disabled Veteran Owned Small Businesses (SDVOSB) or Veteran Owned Small Businesses (VOSB) has the capacity to provide this service. The North American Industry Classification System (NAICS) code for this procurement is 541711, title Research and Development , with a size standard of $15.0 million. Other related work includes: The contractor shall provide all labor, consultation with genetics experts, detail information regarding step-by-step generation of the mice model including sequencing information or others related to genetic alternation made during the process, and other resources required to deliver confirmed F1 mice to VA, guided by the following aims: 1) project strategy 2) molecular engineering for Sptbn1 and Tgfbr2 3) in vivo development and confirmation of founder animals, and 4) founder confirmation of F1 animals, except as may otherwise be specified in this statement of work. 1. Project Strategy Service includes: Consultation with Molecular and Genetics Experts Vetting potential strategies to meet project goals Bioinformatic analysis of the gene of interest 2. Molecular Engineering of Sptbn1 (Gene ID: 20742) and Tgfbr2 (Gene ID: 21813) Service includes: Design of genetic modification strategy in accordance with gene features and model requirements Validation of selected strategy with customer Design and synthesis of CRISPR/Cas9 reagents ESC targeting and pre-screening In vitro validation of CRISPR/Cas9 reagents ESCs used are mixed C57BL/6;129SVjae The objective of this step is to design, generate and deliver reagents enabling generation of a Sptbn1-CREER and Tgfbr2-CREER knockin/out strain. In addition, screening procedures for efficient detection of genetic modification will be designed and set up. 3: in vivo Development Service includes: Preparation of C57BL/6NCrl mouse embryos Preparation of pseudo-pregnant mice (health status VAF/Elite®) Injection into embryos Reimplantation of injected embryos into recipient female mice The objective of this step is to generate potential F0 transgenic mice assuming the genetic modification is not lethal or toxic. 4: Founder Confirmation of F1 animals, except as may otherwise be specified in this statement of work. Service includes: Maintenance and weaning of the mice Breeding of up to 5 F0 transgenic mice with wild type C57BL/6N mice Confirmation of germline transmission and allelic segregation Screening of F1 animals (up to 40 mice) The delivery of animals will take place after genetic characterization and VAF/Elite® health status (approximate age 4-6 weeks). 3. Background: Sptbn1 has two isoforms using completely different exons for start codon and stop codon. To effectively target this, our first choice is to disrupt both isoforms by inserting Cre-ERT-STOP into the shared exon 2 for both isoforms, because the additional AA may interfere with CRE recombinase activity. If this is the case, we will target isoform1 that is more abundant by inserting CRE-ERT-STOP to fist start codon of isoform1. We will choose ESC cell injection that allows the pre-screen of the right ESCs prior to blastocyte injection to generate the founder. 4. Performance Period: Timeline: Experiment design, initiation to ESCs cells targeting 12-16 weeks F0 Founder mice and validation 6-8 weeks F1 confirmation 8-10 weeks The contractor s work shall begin upon initiation (Milestone 1). Milestone 2, the validation of CRISPR reagents ready for injection, must be completed within a 12-16 week time period, Milestone 3, live birth of founder animals and confirmation of F0, must be completed within a 6-8 week time period, and Milestone 4, confirmation of F1 animals, must be completed within an 8-10 week time period for the agreed upon fixed price. To ensure these timelines are met, the contractor will work as per the Government s hours of operation, 9 AM 5 PM from Monday Friday (40-hour workweek). The contractor will not be required to work when the Government is closed for Government holidays. The Government anticipates making a single award of Firm-Fixed price award for this requirement. Disclaimer: This is a Sources Sought only. It does not constitute a Request for Proposal, Invitation for Bid, or Request for Quote and it is not to be construed as a commitment by the Government to enter into a Contract. No solicitation exist at this time; If a solicitation is released it will be synopsized in the Federal Business Opportunities Website, (https://www.fbo.gov). All Contractors must be registered in System Award Management data base, (www.sam.gov). Registration is free and potential offerors are encouraged to visit the SAM website and register as soon as possible to avoid delays. Questions concerning this Sources Sought announcement should be directed to Carla G. Maria, Contract Specialist via email at contact@example.gov.