10 mL of blood is collected from the patient. PBMCs (Peripheral Blood Mononuclear Cell) are further separated from the rest of the blood. These PBMCs will undergo reprogramming by adding the 4 Yamanaka factors OCT4, SOX2, KLF4, C-MYC via viral transvection. The addition of these 4 factors cause the cells to revert back to a stem fate. 2-3 weeks later single cells inducible Pluripotent Stem Cells (iPSC) will be screened for intact Chromosome structure (karyotyping) and tested for their ability to further differentiate. Once iPSC clone screening has identified healthy clones, these clones will be expanded and placed in cryogenic storage for subsequent procedures, or delivery to partner labs.

CRISPR is a cutting-edge high-procession genetic-engineering-tool that allows for addition, deletion, and direct alteration of single DNA-base-pairs within the genomic sequence. CRISPR will be used to make patient iPSCs with the correct DNA sequence. This corrected isogenic iPSC wild-type (WT) cell line will be used as a healthy WT control to patient cell lines allowing demonstration of effect of Anti-Sense-Oligo ASO or small molecules through screenings. CRISPR corrected iPSC clones will be sequenced to demonstrate no other alterations were made to the genomic sequence. Healthy clones will be expanded and placed in cryogenic storage for subsequent procedures, or delivery to partner labs.

As part of a feasibility/amenability study, patient DNA will be sequenced to find the mutation effecting the patient (phenotype). An expert will analyze the genomic data against the scientific literature and prepare a report detailing the next steps for the patient.

As a service we also work with various hospitals, biotechs, and labs who need to sequence DNA or RNA.

iPSC cells along with isogenic controls will be grown up in hundreds of tissue culture plates, differentiated into effected tissues or organoid bodies, and used to run lab tests (assay) developed to see a proper function of a cell restored. ASOs designed to antagonize expression of a deleterious message. Similarly, and an array of small molecule drugs can be tested Each ASO or small molecule will be plated multiple times at multiple concentrations to confirm any positive result. Speed and procession are necessary as potentially thousands of ASO or small molecules may be screened in this fashion.

In the case that the mutation has a dominant and deleterious, dominant-negative, or gain of function, effect it may be necessary to take an allele specific ASO approach to knockdown the expression of a gene. A requirement of this approach is a long read genomic DNA sequencing with greater depth than the first. This is necessary to Phase the mutation to the allele effected. Minor sequences called polymorphisms will be used to target the ASO knockdown to the mutation effected allele.
As a service we also work with various hospitals, biotechs, and labs who need to sequence DNA or RNA.

In cases where speed is the first priority, a skin punch biopsy can be taken from the patient. From this skin punch Fibroblast cells will be isolated and cultured. With some big detractors*, these cells can be used to shortlist ASO or small molecules while work to establish and differentiate iPSCs is being done. *While patient derived fibroblasts cultures are faster to establish they are known to frequently have mutations and continue to mutate during culture. These mutations may effect the outcome of the experiments and are therefore not sufficient for final clinical decisions.

10 mL of blood is collected from the patient. PBMCs (Peripheral Blood Mononuclear Cell) are further separated from the rest of the blood. These PBMCs will undergo reprogramming by adding the 4 Yamanaka factors OCT4, SOX2, KLF4, C-MYC via viral transvection. The addition of these 4 factors cause the cells to revert back to a stem fate. 2-3 weeks later single cells inducible Pluripotent Stem Cells (iPSC) will be screened for intact Chromosome structure (karyotyping) and tested for their ability to further differentiate. Once iPSC clone screening has identified healthy clones, these clones will be expanded and placed in cryogenic storage for subsequent procedures, or delivery to partner labs.

An in-depth understand of how a disease state effects other RNAs and proteins that the malfunction gene/protein interacts with must be developed in order to test if a given treatment is having an effect. Assay development coordinates these studies in an effort to find 1, or a series of lab tests that are fast and easy to run thousands of times in a high throughput fashion.