United States - Small Molecule Modulators of iPSC-Derived Hepatocyte and Neuronal Calcium Signaling

Description

Added: Jul 30, 2019 4:09 pm

SOURCES SOUGHT NOTICE
1. Sources Sought Number: 75N95019Q00282
2. Title: Small Molecule Modulators of iPSC-Derived Hepatocyte and Neuronal Calcium Signaling
3. Classification Code: 99- Miscellaneous
4. NAICS Code: 541714 - Research and Development in Biotechnology (except Nanobiotechnology)
5. Description:
This is a Sources Sought notice. This is NOT a solicitation for proposals, proposal abstracts, or quotations. The purpose of this notice is to obtain information regarding the availability and capability of all qualified sources to perform a potential requirement.
This notice is issued to help determine the availability of qualified companies technically capable of meeting the Government requirement and to determine the method of acquisition. It is not to be construed as a commitment by the Government to issue a solicitation or ultimately award a contract. Responses will not be considered as proposals or quotes. No award will be made as a result of this notice. The Government will NOT be responsible for any costs incurred by the respondents to this notice. This notice is strictly for research and information purposes only.
6. Statement of Need and Purpose: The National Center for Advancing Translational Science (NCATS) of the National Institutes of Health is in need of CRISPR/cell cloning to generate two custom iPSC reporter cell lines for small molecule screening projects. These cell lines include: (1) a custom dual gene knock-in ALB-2A-secNLuc and AFP-2A-eGFP iPSC reporter cell line encoding the aforementioned reporter sequences at their endogenous genomic loci; (2) a custom gene knock-in GCaMP iPSC reporter cell line encoding a GCaMP calcium indicator into a genomic "Safe Harbor" locus. The parental iPSC cell line will be provided by the contractor. These cell lines are critical for identifying chemical modulators of (1) iPSC-derived hepatocyte differentiation and maturation and (2) calcium signaling in iPSC-differentiated neurons. It is a priority for NCATS to discover, validate, and disseminate small molecule reagents to replace expensive recombinant proteins, xenogenic material and undefined media components in cell differentiation protocols.
7. Purpose and Objectives: NCATS' workflow for these projects, the foundation of which is built upon construction of these iPSC reporter cell lines, consists of identifying small molecule modulators of (1) iPSC-derived hepatocyte differentiation and maturation and (2) calcium signaling in iPSC-differentiated neurons from high throughput screening tens of thousands of compounds. Upon retesting in primary and secondary assays, confirmed hits may be selected for optimization through medicinal chemistry. Additional studies would be performed to help identify or characterized the suspected targets of these molecules, and further focus characterization of their activity.
8. Project requirements: This contract should provide CRISPR-mediated genome editing for the following projects in iPSC cell lines: (1) a custom dual gene knock-in ALB-2A-secNLuc and AFP-2A-eGFP iPSC reporter cell line encoding the aforementioned reporter sequences at their endogenous genomic loci; (2) a custom gene knock-in GCaMP iPSC reporter cell line encoding a GCaMP calcium indicator into a genomic "Safe Harbor" locus. The vendor should provide two homozygous clones for each project/cell line containing the indicated reporter knock-ins, 2 vials of each clone with 1 x 106 cells/vial. The vendor should also provide milestone updates/reports, as well as a final report with detailed descriptions of each procedure, including targeting vector design, construction and validation, transfection condition, genotyping strategy, and results. Ownership of these cell lines should be exclusive to NCATS SCTL.
Project 1:
Technology Utilized: CRISPR
Gene Name of Knock-In Target #1: ALB (human albumin)
Gene ID of Knock-In Target #1: ENSG00000163631
Gene Name of Knock-In Target #2: AFP (human alpha fetoprotein)
Gene ID of Knock-In Target #2: ENSG00000081051
Intended Modifications for Target #1: Directly after the endogenous ALB gene, knock in a 2A "slipstreaming" sequence followed by secreted NanoLuc, with the intention of having endogenous ALB and secreted NanoLuc expression controlled by the same ALB endogenous promoter.
Intended Modifications for Target #2: Directly after the endogenous AFP gene, knock in a 2A "slipstreaming" sequence followed by eGFP (or brighter monomeric derivative), with the intention of having endogenous ALB and eGFP expression controlled by the same AFP endogenous promoter.
Project 2:
Technology Utilized: CRISPR
Gene Name of Knock-In: Next-generation GCaMP genetically encoded calcium indicator; specific GCaMP indicator TBD
Gene ID of Knock-In: N/A; gene is synthetic
Intended Modification: Knock in a next-generation GCaMP genetically encoded calcium indicator into a genomic "Safe Harbor" locus

Anticipated period of performance: NCATS expects to issue a purchase order in FY19 (no later than September 30, 2019).
The stages of this contract work will be delivered within the following timeline milestones:
Project 1: Dual gene knock-in ALB-2A-secNLuc and AFP-2A-eGFP iPSC reporter cell line
- ALB-2A-secNLuc gene knock-in
a. iPSC cell line purchase and sequencing of targeted regions: 2-3 weeks
b. CRISPR vector construction: 2-3 weeks
c. Reagent validation and donor plasmid construction: 4-6 weeks
d. Transfection of targeting vectors: 2-4 weeks
e. Cell confirmation and expansion: 3-4 weeks
- AFP-2A-eGFP gene knock-in
a. iPSC cell line purchase and sequencing of targeted regions: 2-3 weeks
b. CRISPR vector construction: 2-3 weeks
c. Reagent validation and donor plasmid construction: 4-6 weeks
d. Transfection of targeting vectors: 2-4 weeks
e. Cell confirmation and expansion: 3-4 weeks
- Project 1 total: 26-40 weeks
Project 2: Gene knock-in GCaMP iPSC reporter cell line
a. iPSC cell line purchase and sequencing of targeted regions: 2-3 weeks
b. CRISPR vector construction: 2-3 weeks
c. Reagent validation and donor plasmid construction: 4-6 weeks
d. Transfection of targeting vectors: 2-4 weeks
e. Cell confirmation and expansion: 3-4 weeks
- Project 2 total: 13-20 weeks
9. Reporting Requirements: Two CRISPR genome-edited iPSC reporter cell lines: (1) a custom dual gene knock-in ALB-2A-secNLuc and AFP-2A-eGFP iPSC reporter cell line encoding the aforementioned reporter sequences at their endogenous genomic loci; (2) a custom gene knock-in GCaMP iPSC reporter cell line encoding a GCaMP calcium indicator into a genomic "Safe Harbor" locus. The details of each project/cell line are indicated under Specific Requirements. Deliverables for each project/cell line include: two homozygous clones containing the indicated reporter knock-ins; 2 vials of each clone with 1 x 106 cells/vial; milestone updates/reports; a final report with detailed descriptions of each procedure, including targeting vector design, construction and validation, transfection condition, genotyping strategy, and results. Ownership of these cell lines should be exclusive to NCATS SCTL.
10. REQUESTED RESPONSE:
The intent of this sources sought notice is to define the procurement strategy (e.g. set-aside, sole source, unrestricted) for a solicitation that NCATS intends to post soon. Interested contractors are requested to respond by (1) identifying how their proposed product meets or exceeds each of the seven salient charactistics listed above, by number; and (2) providing product literature, white papers, or capability statements that demonstrate their bona fide ability to supply such an instrument within the time frame described above.
In addition, please state your business size status and whether you are the manufacturer or authorized distributor of the device you propose.
Please note that NCATS is particularly interested in small business sources. If you are a small business, please state the type of small business (small business; HUBZone; service-disabled, veteran-owned; 8(a); veteran-owned; woman-owned; or small disadvantaged businesses) and your size classification relative to the North American Industry Classification System (NAICS) code for the proposed acquisition. Your responses to the information requested will assist the Government in determining the appropriate acquisition method, including whether a set-aside is possible.
Please note that if no responses to this notice are received from either authorized distributors or providers of potentially equivalent items, then this action will be sole-sourced to the provider cited above.
Please provide your DUNS number, organization name, address, point of contact, other information described above, and any other information that may be helpful in developing or finalizing the acquisition requirements.
The information submitted must be must be in an outline format that addresses each of the elements of the project requirement. A cover page and an executive summary may be included but is not required.
The response must include the respondents' technical and administrative points of contact, including names, titles, addresses, telephone and fax numbers, and e-mail addresses.

11. All responses to this notice must reference number 75N95019Q00282 and be submitted by email to Jessica Adams, Contract Specialist at jessica.adams@nih.gov.
Responses must be received on or before August 2, 2019, at 5:00pm. Eastern Standard time.
12. Disclaimer and Important Notes: This notice does not obligate the Government to award a contract or otherwise pay for the information provided in response. The Government reserves the right to use information provided by respondents for any purpose deemed necessary and legally appropriate. Any organization responding to this notice should ensure that its response is complete and sufficiently detailed to allow the Government to determine the organization's qualifications to perform the work.
Respondents are advised that the Government is under no obligation to acknowledge receipt of the information received or provide feedback to respondents with respect to any information submitted. After a review of the responses received, a presolicitation synopsis and solicitation may be published in Federal Business Opportunities. However, responses to this notice will not be considered adequate responses to a solicitation.
Confidentiality: No proprietary, classified, confidential, or sensitive information should be included in your response. The Government reserves the right to use any non-proprietary technical information in any resultant solicitation(s).
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Opportunity closing date

02 August 2019

Value of contract

to be confirmed