D. Attachment 1 - Statement of Work.pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

STATEMENT OF WORK (SERVICES)

(SOW)

GENERAL INFORMATION

Title of Project:

Custom Conditional Gpnmb cDNA cassette Knockin mouse model

Statement of Need and Purpose:

The purpose of this purchase is for the generation of a Custom Conditional Gpnmb cDNA cassette Knockin mouse model which is required to elucidate the role of Gpnmb specifically in macrophages during skeletal muscle injury responses and aging.

Background Information and Objective:

Macrophages (MΦs) are a heterogenous population of and immune cells that play an essential role in SKM (skeletal muscle) growth repair. Distinct subpopulations of MΦs perform diverse specialized functions. Our lab recently identified 11 distinct MΦ subpopulations in aging mouse SKM by single cell (sc)RNA-sequencing. Of particular interest, cluster 6 (Cl6) increased from 2.5% of the total MΦ population in young mice to 13% in old mice. This subpopulation of MΦs was enriched in mRNAs encoding candidate senescence proteins including GPNMB, SPP1, CTSD and GDF15. We hypothesize that GPNMB+ MΦs represent senescent SKM macrophages that may contribute to age-related SKM phenotypes. We propose to study the role of GPNMB+ MΦ cluster (Cl6), rather than GPNMB itself, in SKM injury response and aging using an inducible GPNMB+ MΦ ablation mouse model. We aim to generate Cre-inducible Gpnmb-diphtheria toxin receptor (DTR) knock-in (KI) mice to enable the specific elimination of GPNMB+ MΦs via DT. We will then elucidate the role of GPNMB+ MΦs in SKM injury repair and SKM aging. This model can be used to analyze the function of GPNMB+ MΦs in other tissues, including the cardiovascular system and brain.

Period of Performance:

Timeline is the estimated time required for order completion, starting from Client approval of targeting strategy proposed by Contractor’s design team. Requests for additional services may alter the actual delivery date. Germline Transmission F1 mice would be delivered to NIA within 9-11 monthsafter receipt of order (ARO).

SCOPE OF WORK

General Requirements:

Independently and not as an agent of the Government, the Contractor shall furnish all the necessary services, qualified personnel, material, equipment, and facilities, not otherwise provided by the Government as needed to perform the Statement of Work below:

Vendor will generate one Conditional DTR cDNA Knockin Mouse Model (Modify mouse Gpnmb gene by inserting DNA cassette containing loxP-STOP-loxP and DTR cDNA at client-specified/TBD insertion site. Genomic target region ≤ 2 Kb, cDNA size ≤ 2 Kb.)

● Deliverable: Germline Transmission F1 Mice

Specific Requirements:

Model Type: Custom Conditional DTR cDNA Knockin Mouse Final Deliverable: Germline-confirmed F1 heterozygous mice Strain: C57BL/6 Gene of interest: Gpnmb Gene ID: ENSMUSG00000029816, ENSMUST00000031840.10 Estimated Timeline: 9-11 months

The vendor will provide monthly updates via email to the government on the project progress including any troubleshooting required.

Live Animal Import: The vendor will provide the required paperwork for the import request when the live animals are ready for shipping to NIA including health reports, facility health program description. These documents will be coordinated between the vendor, the Program Contact, to be identified at time of award, NIA Comparative Medicine Section and the Animal Care and Use Committee as necessary.

LEVEL OF EFFORT:

Contractors field service application specialists or equivalent will spend approximately 9-11 months on generating targeting materials, screening ES cell clones, then additional breeding services as required for Germline-confirmed F1 heterozygous mice. Germline transmission is guaranteed once positive ES cells are identified and injection and breeding will continue as long as it takes to achieve germline transmission.

GOVERNMENT RESPONSIBILITIES

The government will not be providing starting materials to the vendor. All necessary reagents are commercially available to the vendor.

The government has submitted Amendment #6 to their animal protcol ASP 476 to request for the addition of a new mouse strain (Gpnmb-DTRfx) to the NIA facility.

Through project planning discussions, the government will help determine the targeting insertion site for the Gpnmb knockin model with the Vendor.

At the conclusion of the project, the government will work with the vendor to coordinate the proper documents required by NIA Comparative Medicine Section and the Animal Care and Use Committee to import the live mice into the NIA facility.

DELIVERY OR DELIVERABLES

Deliverable: Germline Transmission F1 Mice generated as Cre-inducible Gpnmb-diphtheria toxin receptor (DTR) knock-in (KI) mice

REPORTING REQUIREMENTS

The vendor will provide monthly reports via email to the government on the project progress

OTHER CONSIDERATIONS

Travel:

The principal place of performance is vendor’s laboratory and vivarium.

Key Personnel:

NA

Information System Security Plan:

Data Rights:

Section 508—Electronic and Information Technology Standards:

Publications and Publicity:

Confidentiality of Information: