D_Statement of Work.pdf

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

STATEMENT OF WORK

GENERAL INFORMATION

Title of Project:

Cloned Oligo Pool Libraries

Statement of Need and Purpose:

The purpose of this requirement is to design pooled oligo libraries and primary tranformant tRNAs that improve on existing tools, resulting in more robust gene knockdown and allowing more compact screens, thus reducing the cost of future experimentations.

The objective of this requirement is to generate pooled oligos for genome-wide dual single guide RNA (sgRNA) clustered regularly interspaced short palindromic repeats (CRISPRi) libraries, and large-scale generation of final plasmid libraries for the purpose of performing CRISPRi survival screens in iPSC differentiated into neurons for the Neurodegenerative Initiative (iNDI) project.

Background Information and Objective:

The National Institutes of Health (NIH) Intramural Center for Alzheimer’s and Related Dementias (CARD) requires custom generation of oligos and/or plasmids for the research of neurodegenerative diseases.

The iPSC Neurodegenerative Disease Initiative (iNDI) project has engineered a set of iPSC lines. Using these engineered lines, the generation of this tool allows for a CRISPRi tRNA library to improve on existing tools.

This will result in a more robust gene knockdown and allowing more compact screens. This entails a multi-step cloning method performed at a large-scale to ensure that the final library contains unique tRNAs included within the oligo pools. This requires a high level of expertise and quality control.

Period of Performance:

The anticipated performance period is July 10, 2023 to August 31, 2023.

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.

Specific Requirements:

The vendor shall generate (4) CRISPRi tRNA oligo libraries, and (4) unique primary transformants for intermediate tRNA libraries. These biologicals will improve the projects’ existing tools being used within the INDI initiative which will result in a more robust gene knockdown and allowing more compact screens, thus reducing the cost of each future experimentation. The generation of the oligo pooled libraries and primary transformants require a multi-step cloning method performed at a large-scale to ensure that the final libraries and transformants contain all of the unique tRNAs. The vendor shall provide the following eight (8) requirements:

1. Cloned Oligo Pool Library, Intermediate tRNA library, 21554 customer-defined dual-guide variants, cloned into customer-supplied vector, 250 ug delivered as purified plasmid DNA.

2. Cloned Oligo Pool Library, Intermediate tRNA library, 21554 customer-defined dual-guide variants, cloned into customer-supplied vector, 250 ug delivered as purified plasmid DNA.

3. Cloned Oligo Pool Library, M6_Scaffold_tRNA_Library, 21554 customer defined dual-guide variants with conserved portions from Intermediate tRNA library, cloned into customer-supplied vector with M6 scaffold dropped in via two step cloning, 250 ug delivered as purified plasmid

DNA.

4. Cloned Oligo Pool Library, iF15_2xU6-bsd-BFP_rmLoxP_Final, 21554 customer-defined dual-guide variants, cloned into customer supplied vector with 2xU6 scaffold dropped in via two step cloning, 250 ug delivered as purified plasmid DNA.

5. Primary transformants for Intermediate tRNA library in TG1 cells as ~10 tubes of glycerol stocks.

6. Primary transformants for 1 CR3_Scaffold_tRNA_Library in TG1 cells as ~10 tubes of glycerol stocks.

7. Primary Transformants for M6_Scaffold_tRNA_Library in TG1 cells as ~10 tubes of glycerol stocks.

8. Primary transformants for iF15_2xU6-bsd-BFP_rmLoxP_Final library in TG1 cells as ~10 tubes of glycerol stocks.

This workflow requires the Government to provide custom vectors or plasmids to the vendor, and then using a large scale CRISPRi method, the vendor shall generate large pooled libraries with custom designed variants. These oligo libraries and primary transformant variants will be delivered to the Government.

LEVEL OF EFFORT:

Vendor staff shall provide the oligo libraries and primary transformants when genetic engineering is completed.

GOVERNMENT RESPONSIBILITIES

The government will provide four customer-supplied vectors to the vendor.

DELIVERY OR DELIVERABLES

For each of the supplied custom vector, vendor shall produce 4 oligo pool libraries and 4 unique primary transformant variants.

REPORTING REQUIREMENTS

The vendor shall provide weekly updates for Government on progress of the project via email.

OTHER CONSIDERATIONS

Travel:

Not Applicable. The principal place of performance is the vendor’s work site.

Key Personnel:

Vendor shall provide a dedicated project manager to the government as a point of contact for the services.

Information System Security Plan:

Not Applicable.

Data Rights:

Section 508—Electronic and Information Technology Standards:

The contractor shall comply with Section 508 of the Rehabilitation Act (29 U.S.C. § 794d), as amended by the Workforce Investment Act of 1998 (P.L. 105-220), August 7, 1998.

Publications and Publicity:

Confidentiality of Information: