1-SOW.pdf

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

STATEMENT OF WORK (SERVICES)

(SOW)

GENERAL INFORMATION

Title of Project:

Development of five different iPS reporter cell lines expressing fluorescent protein or HiBiT at specific target genes.

Statement of Need and Purpose:

The SCTL requires the services of Synthego to meet project milestones. Synthego’s services portfolio includes a large selection of gene editing services that allow for the knock-in of reporter tags at specific gene regions in iPS cells. The services offered that are of interest to the SCTL include:

• CRISPR-Edited iPS Cells – using a robust, automated CRISPR editing process that delivers consistently high editing in virtually any cell type. Synthego has over 60,000 edits completed and characterized. This expertise is applied to iPS cell editing to achieve unprecedented editing efficiencies of the highest quality. They offer knockouts, single nucleotide variants, and tags in user-supplied iPS cell lines available in homozygous or heterozygous clone or pool format.

These services will be utilized by the SCTL to obtain reporter cell lines that will facilitate the development of novel iPSC differentiation protocols. The data obtained from this study will be used to generate new iPSC-derived cell types.

Background Information and Objective:

The SCTL has recently developed a protocol for differentiating hPSC into various cell types such as neurons, nociceptors, astrocytes and placental cells. However, one limitation in the field is that many iPSC-derived cell types retain immature phenotypes. Therefore, the development of reporter cell lines for mature gene markers would facilitate the obtention of cell types that more closely resemble their somatic counterparts.

Likewise, the stem cell field has not been able to develop robust protocols for the differentiation of iPSCs to more difficult-to-generate cell types, such as pancreatic beta cells. Here, the proposed reporter cell line set follows the developmental trajectory of stem cells to somatic cell types that would facilitate the establishment of novel differentiation protocols for pancreatic beta cells.

Period of Performance:

The period of performance will commence within 30 days after receipt of order (ARO) and last for about 16 to 25 weeks, depending on the type of tag.

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:

The SCTL will provide the parental cell line. Then, the vendor will QC the cells upon arrival, preform gene editing at specified targets, validate tag knock-in, generate single-cell clones, expand, re-validate knock-in and QC cells. Finally, the vendor will ship the gene-edited iPSCs to

SCTL.

Specific Requirements:

The SCTL will provide the LiPSC G.R. 1.1 parental iPS cell line for gene editing, then the vendor will:

For CRISPR gene editing:

• Qualify cells prior to gene editing

• Design CRISPR quide RNA

• Design donor template for specified genes with HiBiT tag

• Perform electroporation

• Perform pool analysis

• Perform single-cell clonal expansion

• Perform clonal analysis

• Qualify cells after gene editing for knock-in at specified sites, pluripotency and karyotype

• Provide cryopreserved reporter cells and QC data to SCTL.

Specified target genes:

- NKX6.1 (GFP-tagged)

- INS (m-Cherry and HiBiT-tagged in different regions and clones)

- PDX1 (HiBiT-tagged)

- ISL1 (HiBiT-tagged)

LEVEL OF EFFORT:

As per their internal standard operating procedures, Synthego will be responsible for carrying out CRISPR gene editing, single-cell clones and QC of gene-edited iPS cells

GOVERNMENT RESPONSIBILITIES

The SCTL will provide the parental iPS cell line for gene editing.

DELIVERY OR DELIVERABLES

Deliverables:

• Cryopreserved single-cell clonal reporter cell lines for specified genes.

• QC data.

Key Personnel:

N/A

Information System Security Plan:

N/A

Data Rights:

Data generated as a result of all services will be the properties of NIH, NCATS, and SCTL.

Section 508—Electronic and Information Technology Standards:

N/A

Publications and Publicity:

The datasets derived by the SCTL from these experiments will be part of scientific publications and they will be made available to the public on the SCTL public-facing internet website. The contractor will not publish any results of the work.

Confidentiality of Information:

The Contractor will keep the results of the contract confidential.