Inactive Total Small Business Set-Aside (FAR 19.5)

The Contractor shall generate midbrain organoids from six different cell lines that are suitable for RNA sequencing and proteomic analysis. These organoids should contain dopaminergic neurons, a major... The Contractor shall generate midbrain organoids from six different cell lines that are suitable for RNA sequencing and proteomic analysis. These organoids should contain dopaminergic neurons, a major subtype in the midbrain. The work will take approximately 4 months. Aim 1: Generate midbrain organoids from iPSCs Task1: Differentiate and characterize midbrain organoids Detailed Tasks: 1. Differentiate iPSCs into 90-day midbrain organoids: 2 NGLY1 mutant lines (593B and 592D); 2 Wild type lines (JB240 and HT729A); 2 592D corrected lines 2. Characterize midbrain organoids at different stages (14-day, 60-day, and 90-day) to verify their identity 3. Flash freeze the organoids for RNA and protein work 4. Ship the organoids to requester (NCATS) and a third party for analysis Expected Outcome and Milestones: • Generate midbrain organoids that are suitable for RNA sequencing and protein analysis. Deliverables: 1. Characterization of midbrain organoids at different stages. 2. 36 tubes of frozen organoids total: 3 tubes for RNA and 3 tubes for protein from each iPSC line. DELIVERY OR DELIVERABLES Data Package The contractor shall deliver to our servicing CROs, 36 tubes of frozen organoids total: 3 tubes for RNA and 3 tubes for protein from each iPSC line. Support The contractor's scientists will be available to consult with scientists from NCATS throughout the project period and after deliverables are sent. Support will be provided by email, phone, or web meeting per the sponsor preference. The contractor shall send an email update to NCATS every other week. This email will include a summary of progress from the previous weeks, any technical difficulties encountered, and a timeline update. REPORTING REQUIREMENTS The contractor shall send an email update to NCATS every other week. This email will include a summary of progress from the previous weeks, any technical difficulties encountered, and a timeline update. Data Rights: All midbrains generated and data generated is sole property of the government. Contractor has no right to any analyzed iPSCs lines provided by TDB nor to any neurons produced from cells provided by TDB. Note: Contractor shall not distribute or sell these cells and assays developed. They will be discarded at the end of 2021 or at the direction of NCATS.