Bids Are Invited For Service For Generation Of Gene Knock-In Porcine Cells 1. Service For Generation Of Gene ... in India

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Bids Are Invited For Service for Generation of gene knock-in porcine cells 1. Service for Generation of gene knock-in porcine cells The work is to generate multiple gene knock-in cells expressing specific genes. Typically, the service requirement is to a. design, screen and construct gRNA, b. culture of suitable cells c. optimization of transfection d. screening of positive cells and validation e. cell expansion and confirmation of knock-in, f. delivery of vector, cells and detailed experimental protocols. 1. Design of suitable guide RNA for the selected genes and locations in chromosome, synthesis for integration with linear vectors. The final circular vector should have multiple cloning sites. Number of genes to be inserted- Two. The service provider should have more than two guide RNAs designed and tested. 2. Two sets of cells (primary cells such as fibroblasts) to 1 be generated for each knock in one with a fluorescent marker and other without marker. 3. Cell culture, transfection and identification of positive clones, followed by their expansion and confirmation of knock-in. 4. The stepwise progress of work and selection of cells, genes etc. to be done in consultation with ICARNRC on Pig. Deliverables 1. Circular plasmid containing specific guide RNA and Cas nuclease inserted customized for expression in mammalian cells- 4 sets (for two genes, one with and other without marker) in suitable bacteria. 2. 5 million cells per gene to be delivered. The cells should be free frombacterial, viral, mycoplasma and chemical contamination. 3. Detailed protocols and experimental details. All items to be delivered at ICARNRC on Pig, Rani, Guwahati, Assam Total Quantity :

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