D. Attachment 1 - Statement of Work.doc

STATEMENT OF WORK (SERVICES)

(SOW)

GENERAL INFORMATION

Title of Project:

Rat modeling service to create Drd3-Dre and Gal-Cre rat models Statement of Need and Purpose:

The National Institute of Mental Health (NIMH Section on Behavioral Neuroscience requires rat modeling service to create Drd3-Dre and Gal-Cre rat models.

Dopamine receptor 3 (D3) is a receptor subtype that has been implicated in a number of neurodegenerative and neuropsychiatric diseases with behavioral symptoms. D3 is largely localized in an amorphous structure in the mesolimbic dopamine and reward system called the Islands of Calleja (ICj). Gaining transgenic access to D3 neurons allows us to investigate the implications of D3 on cognition and behaviors seen in serious neuropsychiatric/degenerative disorders as well as in the healthy brain via the ICj where D3 may influence brain-wide dopamine release. This has been accomplished in mice but not yet in rats, who are better suited for various measures of cognition than mice.

By using these genetically modified rats, we hope to propose a mechanism of how galanin receptors in the medial septum influence behavior. The understanding of such mechanism can ultimately lead to new ideas about targeted therapeutics that can help improve the quality of life of people who suffer from various mental health disorders.

Background Information and Objective:

The use of D3-cre and GalR1-cre rats are needed to continue a project from previous work in the lab investigating reward-seeking behaviors and dopamine dynamics. It also follows from the long-term goal of the SBN to understand the various components of cognitive-behavioral deficits via the rat. Interest in D3 specifically came about after several recent papers investigating ICj found profound behavioral results from either perturbation or activation. This indicated that this region may be involved in several of the behaviors that we have previously characterized and investigated in rats. However, D3-specific contributions to these behaviors are difficult to investigate as there are many subtypes of dopamine receptors. Access through a transgenic line is therefore the only way to confidently answer many questions related to D3 specifically. Therefore, we determined that the best path forward in the rat, as it has been in mice, is to create a D3-Cre rat. This project follows up from a prior interest of the lab in the neuropeptide galanin. The lab has previously studied the effect of galanin in the hippocampus and prefrontal cortex. The lab now wants to look at the circuitry and function of this neuropeptide in more detail by using genetically modified rats.

Period of Performance:

Estimated timeline is 26 weeks from receipt of order.

SCOPE OF WORK

General Requirements:

The Contractor will create a new transgenic line of long-evans rats targeting the dopamine receptor 3 or galanin 1 gene. This will allow the client access to only neurons that have this receptor subtype present, thus isolating its effects on cognition and behavior. This service is essential in providing an unparalleled level of targeting that cannot be accomplished via other means (e.g. pharmacology, viruses).

Specific Requirements:

· Complete requirements within 26 weeks from receipt of order.

· The Contractor will:

· Provide rat modeling service to create two separate rat models:

· Drd3-Cre rat model

· Gal-Cre rat model

· Design and synthesize an AAV-packaged DNA targeting construct designed to integrate a Cre-P2A construct into the ATG start site of either rat Drd3 and/or Gal.

· Design and synthesize CRISPR sgRNAs targeting these regions of the rat Drd3 and Gal genes for CRISPR-mediated knock-in.

· Create Founder rats via transduction of the AAV-packaged DNA repair template and subsequent electroporation of the CRISPR reagents into Long Evans (Crl:LE) zygotes.

· Will transfer manipulated embryos into surrogate females for gestation and natural delivery.

Requirements and timeline:

Project oversight and labor (includes genotyping assay design)

Project oversight and labor (includes genotyping assay design)

LEVEL OF EFFORT:

· Estimated timelines are provided depending of the efficiency of the genetic engineering process.

· Six stages are estimated for this requirement which must be completed within 24-26 weeks.

GOVERNMENT RESPONSIBILITIES

· The Government will provide the Contractor means to contact NIMH staff so the Contractor can make the necessary updates.

· Government must make shipping arrangements with the Contractor.

DELIVERY OR DELIVERABLES

· Contractor will assist in arranging the live animal shipments, but shipping costs are not included. Contractor must make shipping arrangements with the Government prior to shipping the live animals. A Government Point of Contact will be appointed at the time of award.

REPORTING REQUIREMENTS

· Contractor must provide updates every 4-6 weeks and when requested.

OTHER CONSIDERATIONS

Travel:

There is no travel required for services.

Information System Security Plan:

N/A Data Rights:

Section 508—Electronic and Information Technology Standards:

No Electronic and Information Technology (EIT) will be developed, maintained or used as part of these services.

Publications and Publicity:

Confidentiality of Information:

No personal information or confidential information from the government will be needed to provide these services.

Version 11-30-2016-1

FAR Part 11—Describing Agency Needs