There is a need for improved methods for determining the diagnosis and prognosis of patients with conditions, including autoimmune disease and cancer. Provided herein are methods for using DNA sequencing to identify personalized biomarkers in patients with autoimmune disease and other conditions. Identified biomarkers can be used to determine the disease state for a subject with an autoimmune disease or other condition.

A method of making a prognosis as to whether a patient having renal cancer is likely to survive in a tumour tissue sample obtained from the patient is provided. The method comprising determining the level of expression for each marker of a panel of markers comprising at least one housekeeping gene selected from the group consisting of ACTB, RPL13A, RPL9, and RPS29 and any combinations thereof and at least one prognostic gene selected from the group consisting of CXCL5, EFNA5, EMCN, G6PC, GFPT2, HIST2H3C, IGFBP1, LAMB3, MMP9, MOCOS, PLG, PRAME, RARRES1, SDPR, SLC6A19, TK1, KDELR3 and TSPAN7 and any combinations thereof, comparing the level of expression of each marker with a predetermined reference level associated with each marker, and determining the differential expression of each marker in the tumour tissue sample based on the expression parameter for each marker to provide a prognosis for renal cancer.

Described herein are embodiments of methods to rationally design CRISPR-Cas system-based therapeutics and therapies based on expression of a DNA-damage response signature in a cell. In some embodiments, the methods include screening a set of CRISPR-Cas systems by expressing each CRISPR-Cas system in a test cell population and modifying one or more target sequences in the test cell population; screening in the test cell population for each CRISPR-Cas system and expression of a DNA-damage response signature; and selecting one or more CRISPR-Cas systems that do not result in expression of a DNA-damage response signature.

The present invention provides a low-frequency mutation enrichment sequencing method for free target DNA in plasma, comprising plasma DNA extraction and library construction, general library TT COLD PCR amplification enrichment, probe enrichment capture, PCR and sequencing of captured products, and positive and negatice double-strand error-correction low-frequency information analysis.

Multiplex immunoassays utilize the differential affinities among the conjugation pairs between the capture ligands and target analytes are proposed. Window magnetic-assisted rapid aptamer selection (window-MARAS) methods for selecting aptamers with desirable affinity toward the target analytes and methods for generating reagents for multiplex immunoassays or multiplex detection in one assay by utilizing the selected aptamers as capture ligands in reagents are described and used to demonstrate the feasibility of multiplex immunoassays based on the differential affinity of conjugation pairs between the capture ligands and target analytes.

The disclosure provides methods and compositions that employ gene editing for the treatment of cancer. Gene editing systems specifically target tumor DNA to introduce an expression cassette with a coding sequence that is expressed by tumor cells as a neoantigen that mark the tumor cells for cell death.

The present invention provides methods, compositions and kits for targeted nucleic acid sequence enrichment in a nucleic acid sample and for high efficiency nucleic acid library generation for next generation sequencing (NGS). Specifically, the methods, compositions and kits provided herein are useful for the production and capture of amplification-ready, target-specific and strand-specific regions of interest from nucleic acid samples containing complex DNA.

Provided herein are method of determining the efficacy of targeted therapy in a subject by detecting changes in levels of cell-free tumor load (cfTL). In some aspects, the efficacy of targeted therapy is determined a very short time after the targeted therapy is administered. Also provided herein are method of determining resistance to a targeted therapy in a subject by detecting changes in levels of cell-free tumor load (cfTL).

A method and system for extracting neoantigens for immunotherapy includes the following steps: step S1: acquiring conventional proteomes of tumor tissue and normal tissue samples; step S2: acquiring nucleotide polymer sequence libraries of the tumor tissue and normal tissue samples and a specific proteome of the tumor tissue sample; step S3: acquiring a plurality of candidate tumor-specific neoantigens based on the conventional and specific proteomes of the tumor tissue sample and molecular human leukocyte antigen (HLA) typing; and step S4: calculating the presence of the plurality of candidate tumor-specific neoantigens in the conventional proteomes and the nucleotide polymer sequence libraries of the tumor tissue and normal tissue samples, and acquiring tumor-specific neoantigens with a multiple of gene expression changes as a filter rule. More tumor-specific neoantigens are discovered using the new method because they are not limited to coding regions and are partly derived from genome noncoding regions (NCRs).

Provided herein, in some embodiments, are methods and compositions for identifying combinations of transcription factors, for example, those involved in cell type conversion processes, such as cell differentiation.