Abstract: Disclosed herein, in certain embodiments, are methods of inducing cell cycle arrest, apoptosis, cell death, or a combination thereof, in a cell or population of cells. Also disclosed herein are methods of treating a disease or condition in an individual in need thereof comprising inducing cell cycle arrest, apoptosis, cell death, or a combination thereof in a population of cells in the individual. The cell or population of cells may comprise a nucleic acid sequence associated with a disease or condition, including an autoimmune disease, cancer, or an infectious disease. The methods described herein generally comprise contacting cells with a CRISPR-associated protein and a guide nucleic acid.

Abstract: A cancer vaccine technology is provided which knocks out expression of cell surface immune checkpoint proteins, to facilitate their processing by immune cells, and optionally by knocking-in the expression of cytokines to boost immune response. Non-replicating tumor cells lacking cell surface CD47 are highly effective immunizing agents against subcutaneous mouse melanoma. Whole-cell vaccines inhibited tumor growth, and immunophenotyping showed a dramatic increase in activated effector cell subsets and M1-type macrophages aided by a significant reduction in the tumor-associated macrophage and myeloid derived suppressor cell compartments. A remarkable downregulation of cell surface CD47 was observed in the tumors that did escape after vaccination with genetically modified cells, suggesting the intricate involvement of CD47 in a prophylactic situation. An effective vaccination strategy to increase tumor-specific immune response in solid tumors is provided to improve the outcome of cancer immunotherapy.