Transmembrane receptor gene editing

Abstract

Provided herein are compositions and methods for eliminating intracellular signaling through specific cell surface receptors as a therapeutic approach for a variety of conditions of pro-inflammatory character. In some aspects, the compositions and methods are for preventing the progression of osteoarthritis and other arthritis and for treating osteoarthritis and other arthritis in mammalian joints. In some aspects, the compositions and methods are for treating or preventing localized pain, inflammation, degeneration, or morphological changes associated with back or spinal conditions or disorders.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 301,033, filed January 19, 2022, U.S. Provisional Patent Application No. 63 / 303,479, filed January 26, 2022, and U.S. Provisional Patent Application No. 63 / 390,222, filed July 18, 2022, the contents of which are incorporated herein by reference in their entirety for all purposes. [Background technology]

[0002] Receptor-ligand interactions are involved in the transmission of various signals across the plasma membrane of receptor-expressing cells. In most cases, circulating ligands bind to specific receptors anchored in the cell membrane (with or without co-receptors), which leads to the transduction of signaling pathways from the cell surface to the interior by different mechanisms, and often through the activity of the receptor's cytoplasmic domain. Such signaling events can then affect various cellular activities, including regulating the expression of various gene products that can further affect cellular function.

[0003] The ubiquity of the receptor-ligand paradigm in cell biology means that many diseases, illnesses, and conditions are caused, in whole or in part, by aberrant or excessive signaling through various cellular receptors, and a variety of approaches are available to address this. Small and large molecules can be used to disrupt receptor-ligand interactions, but problems of off-target effects and potential immunogenicity remain.

[0004] More recently, genetic approaches have been explored to either transiently reduce (i.e., knockdown) or permanently eliminate (i.e., gene knockout) the expression of a given ligand or receptor, for example, with siRNA. In both cases, such reduction or elimination can result in "leakyness." For example, a low-affinity receptor may bind to an overabundant ligand in the absence of its natural receptor, or conversely, an unoccupied receptor may bind to other ligands in the absence of its natural ligand. In either case, this leakiness can pose additional problems at the organismal level. Thus, more advanced genetic tools are needed to address the myriad diseases, illnesses, and conditions without the leakiness observed with traditional gene knockdowns or knockouts. Summary of the Invention

[0005] Provided herein are compositions and methods for silencing the signaling function of one or more cellular receptors in an animal in need thereof to treat a disease, illness, or condition caused by aberrant or excessive signaling through the receptor.

[0006] In some embodiments, receptor signaling is silenced by CRISPR editing of the gene encoding the receptor.In some embodiments, CRISPR editing results in the removal of transmembrane domain (for example, the generation of soluble decoy receptor).In some embodiments, CRISPR editing results in the removal of cytoplasmic domain (for example, the generation of membrane-bound decoy receptor).In some embodiments, CRISPR editing results in the targeting of extracellular domain (for example, the generation of complete knockout).

[0007] In contrast to complete ablation or transient knockdown of receptor or ligand gene expression, which are associated with potentially significant off-target effects and leakiness, the targeting of the transmembrane or cytoplasmic domain of a receptor described herein comprises a novel approach to block intracellular signaling of one or more receptor-ligand interactions without the problems of excess ligand or unoccupied receptors contributing to more adverse outcomes. [Brief explanation of the drawings]

[0008] Embodiments of the present disclosure will be further described with reference to the accompanying drawings. The drawings shown are not necessarily to scale, emphasis instead generally being placed upon illustrating the principles of embodiments of the present disclosure.

[0009] [Figure 1] 1 shows exons that can be edited by the CRISPR methods described herein to generate a directed interference (e.g., gene knockout, soluble decoy receptor, or membrane-bound decoy receptor) for a designated exemplary gene target. [Figure 2A] (A) Different types of CRISPR editing are shown for the canine IL1RAP gene and (B) the human TNFRSF4 gene. For cIL1RAP, the top splicing map shows which exons are edited by the indicated sgRNAs and aligned with the corresponding domains in the full-length cIL1RAP protein, with OCP02 editing in the ectodomain, OCP07 editing in the transmembrane domain, and OCP10 editing in the TIR domain (bottom). For TNFRSF4, the bottom splicing map labels the encoded protein domains and aligns with the sgRNA binding sites and generated interference forms. For example, those sgRNAs that generate soluble decoy receptors cluster within the region encoding the transmembrane domain, while those that generate transmembrane decoy receptors cluster within the encoded cytoplasmic domain, eliminating the TRAF binding site. In both cases, ectodomain synthesis is unaffected, allowing ligand binding. [Figure 2B](A) Different types of CRISPR editing are shown for the canine IL1RAP gene and (B) the human TNFRSF4 gene. For cIL1RAP, the top splicing map shows which exons are edited by the indicated sgRNAs and aligned with the corresponding domains in the full-length cIL1RAP protein, with OCP02 editing in the ectodomain, OCP07 editing in the transmembrane domain, and OCP10 editing in the TIR domain (bottom). For TNFRSF4, the bottom splicing map labels the encoded protein domains and aligns with the sgRNA binding sites and generated interference forms. For example, those sgRNAs that generate soluble decoy receptors cluster within the region encoding the transmembrane domain, while those that generate transmembrane decoy receptors cluster within the encoded cytoplasmic domain, eliminating the TRAF binding site. In both cases, ectodomain synthesis is unaffected, allowing ligand binding. [Figure 3A] A non-exhaustive list of disease states associated with IL1R1 and IL1RAP activity is provided. [Figure 3B] A non-exhaustive list of disease states associated with IL1R1 and IL1RAP activity is provided. [Figure 4A] A non-exhaustive list of disease states associated with (A) IL6R and (B) IL6ST activity is shown. [Figure 4B] A non-exhaustive list of disease states associated with (A) IL6R and (B) IL6ST activity is shown. [Figure 5] A non-exhaustive list of disease states associated with TNFRSF1A activity is provided. [Figure 6] A non-exhaustive list of disease states associated with TNFRSF1B activity is provided. [Figure 7] A non-exhaustive list of disease states associated with TNFRSF3 activity is provided. [Figure 8] A non-exhaustive list of disease states associated with TNFRSF4 activity is provided. [Figure 9] A non-exhaustive list of disease states associated with TNFRSF11A activity is provided. [Figure 10A] A non-exhaustive list of disease states associated with (A) TGFBR1 and (B) TGFBR2 activity is shown. [Figure 10B] A non-exhaustive list of disease states associated with (A) TGFBR1 and (B) TGFBR2 activity is shown. [Figure 11A] 1 shows an exemplary sgRNA design targeting canine IL1R1, including (A) an overview of sgRNAs selected based on off-target risk, on-target efficacy, and frameshift potential, and (B) AlphaFold2 models of wild-type and decoy IL1R1 receptors predicted to be generated by OCR13 and OCR14. [Figure 11B] 1 shows an exemplary sgRNA design targeting canine IL1R1, including (A) an overview of sgRNAs selected based on off-target risk, on-target efficacy, and frameshift potential, and (B) AlphaFold2 models of wild-type and decoy IL1R1 receptors predicted to be generated by OCR13 and OCR14. [Figure 12] Figure 1 shows the in vitro performance of tested sgRNA candidates targeting canine IL1RAP as deduced from Sanger traces. ND, not determined. [Figure 13A] 1 shows the effect of different Cas9 variants on the in vitro editing performance of selected candidate sgRNAs targeting canine IL1R1. [Figure 13B] 1 shows the effect of different Cas9 variants on the in vitro editing performance of selected candidate sgRNAs targeting canine IL1R1. [Figure 13C] 1 shows the effect of different Cas9 variants on the in vitro editing performance of selected candidate sgRNAs targeting canine IL1R1. [Figure 13D] 1 shows the effect of different Cas9 variants on the in vitro editing performance of selected candidate sgRNAs targeting canine IL1R1. [Figure 14A]Illustrates the design of an exemplary sgRNA targeting canine IL1RAP, including (A) an overview of sgRNAs selected based on off-target risk, on-target efficiency, and frameshift potential, and (B) AlphaFold2 predicted models of the 3D structures of normal IL1RAP and OCP07-edited IL1RAP. [Figure 14B] Illustrates the design of an exemplary sgRNA targeting canine IL1RAP, including (A) an overview of sgRNAs selected based on off-target risk, on-target efficiency, and frameshift potential, and (B) AlphaFold2 predicted models of the 3D structures of normal IL1RAP and OCP07-edited IL1RAP. [Figure 15] 1 shows the in vitro performance of tested sgRNA candidates targeting canine IL1RAP as deduced from Sanger traces. [Figure 16A] (A) Selected sgRNAs targeting IL1RAP for testing and their editing efficacies with wild-type Cas9, (B) the effect of the indicated Cas9 variants on editing efficacies in canine monocytes, and (C) a comparison of editing efficacies between AR-Cas9 and WT-Cas9 in canine synovial fibroblasts. [Figure 16B] (A) Selected sgRNAs targeting IL1RAP for testing and their editing efficacies with wild-type Cas9, (B) the effect of the indicated Cas9 variants on editing efficacies in canine monocytes, and (C) a comparison of editing efficacies between AR-Cas9 and WT-Cas9 in canine synovial fibroblasts. [Figure 16C] (A) Selected sgRNAs targeting IL1RAP for testing and their editing efficacies with wild-type Cas9, (B) the effect of the indicated Cas9 variants on editing efficacies in canine monocytes, and (C) a comparison of editing efficacies between AR-Cas9 and WT-Cas9 in canine synovial fibroblasts. [Figure 17A] (A) Editing efficacy of the indicated IL1RAP-directed sgRNAs in canine monocytes, (B) canine chondrocytes, and (C) canine synovial fibroblasts. [Figure 17B] (A) Editing efficacy of the indicated IL1RAP-directed sgRNAs in canine monocytes, (B) canine chondrocytes, and (C) canine synovial fibroblasts. [Figure 17C] (A) Editing efficacy of the indicated IL1RAP-directed sgRNAs in canine monocytes, (B) canine chondrocytes, and (C) canine synovial fibroblasts. [Figure 18] 1 shows the effect of IL1R1 editing on silencing the transcriptional induction of PTGS2 in response to 4 hours of exposure to IL1β in canine synoviocytes. [Figure 19] 1 shows the effect of IL1R1 editing on silencing the transcriptional induction of IL6 in response to 4 hours of exposure to IL1β in canine synoviocytes. [Figure 20] We present the experimental design underlying transcriptome analysis, among other assays. Overall, IL1β treatment induces pro-inflammatory signaling solely through the IL1 receptor, whereas MSU crystals and LPS induce inflammation by different means (in addition to inducing IL1 signaling). [Figure 21A] Figure 1 shows transcriptional upregulation of PTGS2 (COX-2) in wild-type control and IL1RAP-edited canine monocytes upon (A) 4-hour and (B) 24-hour exposure to LPS (1 ug / ml), IL1β (100 pM), MSU crystals (400 ug / ml), and PBS (1X). [Figure 21B] Figure 1 shows transcriptional upregulation of PTGS2 (COX-2) in wild-type control and IL1RAP-edited canine monocytes upon (A) 4-hour and (B) 24-hour exposure to LPS (1 ug / ml), IL1β (100 pM), MSU crystals (400 ug / ml), and PBS (1X). [Figure 22A] 1 shows the effect of IL1RAP editing on silencing transcriptional induction of PTGS2 in (A) canine chondrocytes and (B) canine synoviocytes in response to 4 hours of exposure to IL1β. [Figure 22B] 1 shows the effect of IL1RAP editing on silencing transcriptional induction of PTGS2 in (A) canine chondrocytes and (B) canine synoviocytes in response to 4 hours of exposure to IL1β. [Figure 23A] Luciferase activity levels (RLU) are shown in canine synovial fibroblasts (A) after 8 hours and in canine chondrocytes (B) after 24 hours of exposure to luciferase mRNA containing LNPs at the indicated concentrations. [Figure 23B] Luciferase activity levels (RLU) are shown in canine synovial fibroblasts (A) after 8 hours and in canine chondrocytes (B) after 24 hours of exposure to luciferase mRNA containing LNPs at the indicated concentrations. [Figure 24A] Results of synovial fluid cytology assays collected at (A) baseline and (B) 4 weeks after injection of saline, low-dose LNP formulation A, and low-dose LNP formulation B are shown. [Figure 24B] Results of synovial fluid cytology assays collected at (A) baseline and (B) 4 weeks after injection of saline, low-dose LNP formulation A, and low-dose LNP formulation B are shown. [Figure 25A] (A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25B] (A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25C](A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25D] (A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25E] (A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25F] (A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25G](A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 25H] (A) Principal component analysis plot of normalized gene counts after IL1β or MSU treatment for control and IL1RAP-edited cells collectively shown. (B) Two heat maps of selected gene expression as a result of IL1β or (C) MSU treatment, as well as Venn diagrams of up- or down-regulated genes after (D, E) IL1β or (F, G) MSU treatment. (H) Transcriptome analysis of the top 500 IL1β-responsive genes in IL1RAP-edited canine monocyte DH82. [Figure 26A] SEQ ID NOs: 680-824, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1R1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 2 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 26B] SEQ ID NOs: 680-824, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1R1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 2 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 26C]SEQ ID NOs: 680-824, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1R1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 2 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 26D] SEQ ID NOs: 680-824, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1R1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 2 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 26E] SEQ ID NOs: 680-824, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1R1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 2 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 26F] SEQ ID NOs: 680-824, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1R1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 2 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27A]SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27B] SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27C] SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27D] SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27E] SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27F] SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27G] SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 27H]SEQ ID NOs: 825-892 and 3336-3420, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1R1 to generate gene knockouts, soluble decoy receptors, or membrane-bound decoy receptors, or other forms; (E-H) additional information includes chromosome 10 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 28A]

[0023] SEQ ID NOS:893-967 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target equine IL1R1 to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the chromosome 15 genomic coordinates of the bound DNA (assembly equCab3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 28B]

[0023] SEQ ID NOS:893-967 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target equine IL1R1 to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the chromosome 15 genomic coordinates of the bound DNA (assembly equCab3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 28C]

[0023] SEQ ID NOS:893-967 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target equine IL1R1 to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the chromosome 15 genomic coordinates of the bound DNA (assembly equCab3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 29A]SEQ ID NOS:968-1039 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target feline IL1R1 to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the A3 chromosome genomic coordinates of the bound DNA (assembly felCat9), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 29B] SEQ ID NOS:968-1039 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target feline IL1R1 to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the A3 chromosome genomic coordinates of the bound DNA (assembly felCat9), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 29C] SEQ ID NOS:968-1039 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target feline IL1R1 to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the A3 chromosome genomic coordinates of the bound DNA (assembly felCat9), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30A] SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30B]SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30C] SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30D] [Figure 30E] SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30F] SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30G]SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 30H] SEQ ID NOs: 1040-1203, (A-D) crRNA sequences generated by the bioinformatic methods described herein targeting human IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 31A] SEQ ID NOs: 1204-1271 and 3421-3490, (A-C) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms; (D-F) additional information includes chromosome 34 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 31B] SEQ ID NOs: 1204-1271 and 3421-3490, (A-C) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms; (D-F) additional information includes chromosome 34 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 31C]SEQ ID NOs: 1204-1271 and 3421-3490, (A-C) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms; (D-F) additional information includes chromosome 34 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 31D] SEQ ID NOs: 1204-1271 and 3421-3490, (A-C) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms; (D-F) additional information includes chromosome 34 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 31E] SEQ ID NOs: 1204-1271 and 3421-3490, (A-C) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms; (D-F) additional information includes chromosome 34 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 31F] SEQ ID NOs: 1204-1271 and 3421-3490, (A-C) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting canine IL1RAP to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms; (D-F) additional information includes chromosome 34 genomic coordinates of the bound DNA (assembly canFam3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 32A] SEQ ID NOs:1272-1348 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target equine IL1RAP to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the chromosome 19 genomic coordinates of the bound DNA (assembly equCab3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 32B] SEQ ID NOs:1272-1348 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target equine IL1RAP to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the chromosome 19 genomic coordinates of the bound DNA (assembly equCab3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 32C] SEQ ID NOs:1272-1348 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target equine IL1RAP to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the chromosome 19 genomic coordinates of the bound DNA (assembly equCab3), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 33A] SEQ ID NOS: 1349-1424 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target feline IL1RAP to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the combined chromosome C2 genomic coordinates (assembly felCat9), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 33B]SEQ ID NOS: 1349-1424 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target feline IL1RAP to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the combined chromosome C2 genomic coordinates (assembly felCat9), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 33C] SEQ ID NOS: 1349-1424 collectively represent crRNA sequences generated by the bioinformatic methods described herein that target feline IL1RAP to generate (A) a gene knockout, (B) a soluble decoy receptor, or (C) a membrane-bound decoy receptor. Additional information includes the combined chromosome C2 genomic coordinates (assembly felCat9), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 34A] SEQ ID NOs: 1425-1546, (A-C) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (D-F) additional information regarding the chromosome 9 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 34B] SEQ ID NOs: 1425-1546, (A-C) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (D-F) additional information regarding the chromosome 9 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 34C]SEQ ID NOs: 1425-1546, (A-C) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (D-F) additional information regarding the chromosome 9 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 34D] SEQ ID NOs: 1425-1546, (A-C) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (D-F) additional information regarding the chromosome 9 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 34E] SEQ ID NOs: 1425-1546, (A-C) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (D-F) additional information regarding the chromosome 9 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 34F] SEQ ID NOs: 1425-1546, (A-C) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR1 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (D-F) additional information regarding the chromosome 9 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35A]SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35B] SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35C] SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35D] [Figure 35E] SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35F]SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35G] SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 35H] SEQ ID NOs: 1547-1745, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TGFBR2 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 3 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36A] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36B]SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36C] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36D] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36E] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36F]SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36G] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36H] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36I] SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 36J]SEQ ID NOs: 1746-1968, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6R to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37A] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37B] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37C] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37D]SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37E] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37F] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37G] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37H]SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37I] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 37J] SEQ ID NOs: 1969-2178, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human IL6ST to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 5 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 38A] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38B]SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38C] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38D] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38E] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38F]SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38G] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38H] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38I] SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 38J]SEQ ID NOs: 2179-2395, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, (F-J) chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and additional information regarding some predicted performance metrics. [Figure 39A] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39B] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39C] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39D]SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39E] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39F] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39G] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39H]SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39I] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 39J] SEQ ID NOs: 2396-2642, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF1B to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40A] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40B]SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40C] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40D] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40E] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40F]SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40G] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40H] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40I] SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 40J]SEQ ID NOs: 2643-2866, (A-E) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF3 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (F-J) additional information regarding the chromosome 12 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41A] SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41B] SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41C] SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41D]SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41E] SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41F] SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41G] SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 41H]SEQ ID NOs: 2867-3041, (A-D) collectively show crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF4 to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (E-H) additional information regarding the chromosome 1 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42A] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42B] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42C] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42D]SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42E] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42F] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42G] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42H]SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42I] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42J] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42K] SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 42L]SEQ ID NOs: 3042-3335, (A-F) crRNA sequences generated by the bioinformatic methods described herein targeting human TNFRSF11A to generate gene knockouts, soluble decoy receptors, membrane-bound decoy receptors, or other forms, and (G-L) additional information regarding the chromosome 18 genomic coordinates of the bound DNA (assembly hg38), the targeted DNA strand, the targeted exon, and several predicted performance metrics. [Figure 43A] (A) Schematic diagram showing the targeting domain of TGFBR1 and the orientation of various sgRNAs predicted to generate knockout, membrane-bound decoy receptor, and soluble decoy receptor (ECD: extracellular domain, TMD: transmembrane domain, ICD: intracellular domain, GSM: GS-rich motif), and (B) a series of parameters considered for the design of sgRNAs against TGFBR1, as well as exemplary crRNA sequences. [Figure 43B] (A) Schematic diagram showing the targeting domain of TGFBR1 and the orientation of various sgRNAs predicted to generate knockout, membrane-bound decoy receptor, and soluble decoy receptor (ECD: extracellular domain, TMD: transmembrane domain, ICD: intracellular domain, GSM: GS-rich motif), and (B) a series of parameters considered for the design of sgRNAs against TGFBR1, as well as exemplary crRNA sequences. [Figure 44A] (A) Schematic diagram showing the targeting domain of TGFBR2 and the orientation of various sgRNAs predicted to generate knockout, membrane-bound decoy receptor, and soluble decoy receptor (ECD: extracellular domain, TMD: transmembrane domain, ICD: intracellular domain, GSM: GS-rich motif), and (B) a series of parameters considered for the design of sgRNAs against TGFBR2, as well as exemplary crRNA sequences. [Figure 44B](A) Schematic diagram showing the targeting domain of TGFBR2 and the orientation of various sgRNAs predicted to generate knockout, membrane-bound decoy receptor, and soluble decoy receptor (ECD: extracellular domain, TMD: transmembrane domain, ICD: intracellular domain, GSM: GS-rich motif), and (B) a series of parameters considered for the design of sgRNAs against TGFBR2, as well as exemplary crRNA sequences. [Figure 45A] A summary of the efficiency for editing the human TGFBR1 gene in THP-1 cells using the identified guides and (A) wild-type SpCas9 or (B) ARCas9. [Figure 45B] A summary of the efficiency for editing the human TGFBR1 gene in THP-1 cells using the identified guides and (A) wild-type SpCas9 or (B) ARCas9. [Figure 46A] A summary of the efficiency for editing the human TGFBR2 gene in THP-1 cells using the identified guides and (A) wild-type SpCas9 or (B) ARCas9. [Figure 46B] A summary of the efficiency for editing the human TGFBR2 gene in THP-1 cells using the identified guides and (A) wild-type SpCas9 or (B) ARCas9. [Figure 47A] Shown are the relative levels of (A, C) TGFB1 and (B, D) TIMP1 gene expression in THP-1 cells that were unedited (WT) or edited to knock out TGFB1 (OHTG), TGFBR1 (OHTIR), or TGFBR2 (OHTIIR) 6 hours after challenge with LPS (panels A and B) or TGFbeta (panels C and D). [Figure 47B] Shown are the relative levels of (A, C) TGFB1 and (B, D) TIMP1 gene expression in THP-1 cells that were unedited (WT) or edited to knock out TGFB1 (OHTG), TGFBR1 (OHTIR), or TGFBR2 (OHTIIR) 6 hours after challenge with LPS (panels A and B) or TGFbeta (panels C and D). [Figure 47C] Shown are the relative levels of (A, C) TGFB1 and (B, D) TIMP1 gene expression in THP-1 cells that were unedited (WT) or edited to knock out TGFB1 (OHTG), TGFBR1 (OHTIR), or TGFBR2 (OHTIIR) 6 hours after challenge with LPS (panels A and B) or TGFbeta (panels C and D). [Figure 47D] Shown are the relative levels of (A, C) TGFB1 and (B, D) TIMP1 gene expression in THP-1 cells that were unedited (WT) or edited to knock out TGFB1 (OHTG), TGFBR1 (OHTIR), or TGFBR2 (OHTIIR) 6 hours after challenge with LPS (panels A and B) or TGFbeta (panels C and D). [Figure 48A] (A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48B](A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48C] (A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48D](A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48E] (A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48F](A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48G] (A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48H](A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48I] (A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48J](A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). [Figure 48K] (A) hIL1R1 (SEQ ID NOs: 3491-3513), (B) hIL1RAP (SEQ ID NOs: 3514-3543), (C) hIL6R (SEQ ID NOs: 3544-3566), (D) hIL6ST (SEQ ID NOs: 3567-3606), (E) hTNFRSF1A (SEQ ID NOs: 3607-3647), and (F) hTNFRSF1B for use with sgRNA to validate in vitro editing with different delivery modes. Collectively shown are human-directed crRNA sequences targeting (SEQ ID NOs: 3648-3692), (G) hTNFRSF3 (SEQ ID NOs: 3693-3713), (H) hTNFRSF4 (SEQ ID NOs: 3714-3740), (I) hTNFRSF11A (SEQ ID NOs: 3741-3788), (J) hTGFBR1 (SEQ ID NOs: 3789-3813), and (K) hTGFBR2 (SEQ ID NOs: 3814-3865). DETAILED DESCRIPTION OF THE INVENTION

[0010] I. Introduction Provided herein are compositions and methods for silencing the signaling function of one or more cellular receptors in an animal in need thereof to treat a disease, illness, or condition caused by aberrant or excessive signaling through the receptor.

[0011] In some embodiments, receptor signal transduction is silenced by CRISPR editing of the gene that encodes receptor.In some embodiments, CRISPR editing causes transmembrane domain removal (i.e., produces soluble decoy receptor).In some embodiments, CRISPR editing causes cytoplasmic domain removal (i.e., produces membrane-bound decoy receptor).

[0012] II. Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications referenced herein are incorporated by reference in their entirety.

[0013] The term "interleukin-1 receptor type 1" or "IL1R1" refers to the gene (NCBI Gene ID: 3554 [human], NCBI Gene ID: 481328 [dog], NCBI Gene ID: 100009699 [horse], NCBI Gene ID: 101080705 [cat]), or the encoded gene product (e.g., UniProt: P14778; NP_001307909.1 [human], XP_038536135.1 [dog], NP_001075263.2 [horse], XP_023107327.2 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Canonically, the proteins encoded by the genes listed above can bind all forms of the pro-inflammatory cytokine interleukin-1 (IL1 or IL1) and mediate interleukin-1-dependent activation of NF-kappa-B, MAPK, and other signaling pathways. This intracellular signaling involves the recruitment of adaptor molecules such as TOLLIP, MYD88, and IRAK1 or IRAK2 via TIR-TIR interactions with the cytoplasmic domains of the receptor / co-receptor subunits. IL1R1 can also bind to interleukin-1 receptor antagonist (IL1Ra or IL1Ra or IL1RN), which prevents its association with IL1RAP to form a signaling-competent complex. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0014] In certain embodiments, any region of the IL1R1 gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, exon 13, exon 14, exon 15, exon 16, exon 17, exon 18, exon 19, exon 20, exon 21, any intervening intronic regions, intron / exon junctions, the 3' UTR, or the polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the IL1R1 gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the IL1R1 gene targeted by the RNA-guided nuclease is derived from a human (hIL1R1). In some embodiments, the IL1R1 gene targeted by the RNA-guided nuclease is of canine origin (cIL1R1). In some embodiments, the IL1R1 gene targeted by the RNA-guided nuclease is of equine origin (eIL1R1). In some embodiments, the IL1R1 gene targeted by the RNA-guided nuclease is of feline origin (fIL1R1).

[0015] The term "interleukin-1 receptor accessory protein," "IL1RAP," or "IL1RAP" refers to the gene (NCBI Gene ID: 3556 [human], NCBI Gene ID: 488126 [dog], NCBI Gene ID: 100068726 [horse], NCBI Gene ID: 101094125 [cat]), or the encoded gene product (e.g., UniProt: Q9NPH3; NP_002173.1 [human], XP_038318680.1 [dog], XP_001498597.2 [horse], XP_044893081.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Canonically, the proteins encoded by the above-listed genes can associate with IL1-bound IL1R1 to form a high-affinity interleukin-1 receptor complex, which mediates interleukin-1-dependent activation of NF-kappa-B and other signaling pathways through recruitment of adaptor molecules such as TOLLIP, MYD88, and IRAK1 or IRAK2 via TIR-TIR interactions with the cytoplasmic domains of the receptor / co-receptor subunits. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to human, canine, equine, and feline forms, respectively).

[0016] In certain embodiments, any region of the IL1RAP gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, exon 13, exon 14, exon 15, exon 16, exon 17, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene (see, e.g., Figure 2A). In some embodiments, the IL1RAP gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the IL1RAP gene targeted by the RNA-guided nuclease is derived from a human (hIL1RAP). In some embodiments, the IL1RAP gene targeted by the RNA-guided nuclease is derived from a dog (cIL1RAP). In some embodiments, the IL1RAP gene targeted by the RNA-guided nuclease is of equine origin (eIL1RAP). In some embodiments, the IL1RAP gene targeted by the RNA-guided nuclease is of feline origin (fIL1RAP).

[0017] The term "interleukin-1 receptor complex" or "IL1R complex" refers to any number of protein receptors comprising a family of transmembrane protein receptors involved in transmitting or regulating intracellular signaling through the binding of the pro-inflammatory cytokine interleukin-1 (IL1). These members include IL1R1, IL1R2, which function primarily as decoy receptors, IL1RAP, and IL1RL1, which can form complexes with other family members in the IL33 or IL36 signaling systems.

[0018] The term "transforming growth factor beta receptor 1" or "TGFBR1" refers to the gene (NCBI Gene ID: 7046 [human], NCBI Gene ID: 481628 [dog], NCBI Gene ID: 100034117 [horse], NCBI Gene ID: 101094057 [cat]), or the encoded gene product (e.g., UniProt: P36897; NP_004603.1 [human], XP_038538191.1 [dog], XP_023485510.1 [horse], XP_023098269.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Canonically, the proteins encoded by the genes listed above are transmembrane serine / threonine kinases that, together with TGFBR2, form the natural receptors for the TGF-beta cytokines TGFB1, TGFB2, and TGFB3. Upon binding to its ligand, TGFBR1 is phosphorylated by TGFBR2 and activates intracellular signaling that controls multiple physiological and pathological processes through the release of SMAD2 (which can then translocate to the nucleus) or the activation of other cytoplasmic signaling mediators. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of protein or gene (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0019] In certain embodiments, any region of the TGFBR1 gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TGFBR1 gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TGFBR1 gene targeted by the RNA-guided nuclease is derived from a human (hTGFBR1). In some embodiments, the TGFBR1 gene targeted by the RNA-guided nuclease is derived from a dog (cTGFBR1). In some embodiments, the TGFBR1 gene targeted by the RNA-guided nuclease is derived from a horse (eTGFBR1). In some embodiments, the TGFBR1 gene targeted by the RNA-guided nuclease is of feline origin (fTGFBR1).

[0020] The term "transforming growth factor beta receptor 2" or "TGFBR2" refers to the gene (NCBI Gene ID: 7048 [human], NCBI Gene ID: 477039 [dog], NCBI Gene ID: 100033860 [horse], NCBI Gene ID: 101091725 [cat]), or the encoded gene product (e.g., UniProt: P37173; NP_003233.4 [human], XP_038288013.1 [dog], XP_023475502.1 [horse], XP_023116415.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Canonically, the proteins encoded by the genes listed above are transmembrane serine / threonine kinases that, together with TGFBR2, form the natural receptors for the TGF-beta cytokines TGFB1, TGFB2, and TGFB3. Upon binding to its ligand, TGFBR1 is phosphorylated by TGFBR2 and activates intracellular signaling that controls multiple physiological and pathological processes through the release of SMAD2, which can then translocate to the nucleus or activate other cytoplasmic signaling mediators. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to human, canine, equine, and feline forms, respectively).

[0021] In certain embodiments, any region of the TGFBR2 gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TGFBR2 gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TGFBR2 gene targeted by the RNA-guided nuclease is derived from a human (hTGFBR2). In some embodiments, the TGFBR2 gene targeted by the RNA-guided nuclease is derived from a dog (cTGFBR2). In some embodiments, the TGFBR2 gene targeted by the RNA-guided nuclease is derived from a horse (eTGFBR2). In some embodiments, the TGFBR2 gene targeted by the RNA-guided nuclease is of feline origin (fTGFBR2).

[0022] The term "interleukin-6 receptor" or "IL6R" refers to a gene (NCBI Gene ID: 3560 [human], NCBI Gene ID: 612271 [dog], NCBI Gene ID: 102148787 [horse], NCBI Gene ID: 101085689 [cat]) or the encoded gene product (e.g., UniProt: P08887; CAA41231.1 [human], XP_038527979.1 [dog], XP_023496854.1 [horse], XP_023103841.2 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Typically, the proteins encoded by the above-listed genes are transmembrane proteins capable of binding to its natural ligand, interleukin-6. This binding event triggers intracellular signaling events that result in a proinflammatory response. See generally Wolf, J., et al. (2014). Cytokine, 70(1), 11-20. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0023] In certain embodiments, any region of the IL6R gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, exon 13, exon 14, exon 15, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the IL6R gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the IL6R gene targeted by the RNA-guided nuclease is derived from a human (hIL6R). In some embodiments, the IL6R gene targeted by the RNA-guided nuclease is derived from a dog (cIL6R). In some embodiments, the IL6R gene targeted by the RNA-guided nuclease is derived from a horse (eIL6R). In some embodiments, the IL6R gene targeted by the RNA-guided nuclease is of feline origin (fIL6R).

[0024] The term "interleukin-6 cytokine family signal transducer," "GP130," or "IL6ST" refers to the gene (NCBI Gene ID: 3572 [human], NCBI Gene ID: 403545 [dog], NCBI Gene ID: 100051700 [horse], NCBI Gene ID: 101089832 [cat]), or the encoded gene product (e.g., UniProt: P40189; NP_001177910.1 [human], NP_001273950.1 [dog], XP_023481030.1 [horse], XP_011281205.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Canonically, the proteins encoded by the genes listed above are signal transducers shared by many cytokines, including interleukin 6 (IL6), ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), and oncostatin M (OSM), and function as part of a cytokine receptor complex. Activation of this protein depends on the binding of cytokines to their receptors (e.g., IL6 to IL6R). Knockout studies in mice suggest that this gene plays an important role in regulating muscle cell apoptosis. See generally Martinez-Perez, C., et al. (2021). Journal of Personalized Medicine, 11(7), 618. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0025] In certain embodiments, any region of the IL6ST gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, exon 13, exon 14, exon 15, exon 16, exon 17, exon 18, exon 19, exon 20, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the IL6ST gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the IL6ST gene targeted by the RNA-guided nuclease is derived from a human (hIL6ST). In some embodiments, the IL6ST gene targeted by the RNA-guided nuclease is derived from a dog (cIL6ST). In some embodiments, the IL6ST gene targeted by the RNA-guided nuclease is of equine origin (eIL6ST). In some embodiments, the IL6ST gene targeted by the RNA-guided nuclease is of feline origin (fIL6ST).

[0026] The term "tumor necrosis factor receptor 1" or "TNFRSF1A" refers to a gene (NCBI Gene ID: 7132 [human], NCBI Gene ID: 403634 [dog], NCBI Gene ID: 100059548 [horse], NCBI Gene ID: 493957 [cat]), or the encoded gene product (e.g., UniProt: P19438; NP_001056.1 [human], XP_038295153.1 [dog], XP_023498787.1 [horse], NP_001009361.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Typically, the proteins encoded by the above-listed genes are transmembrane receptor proteins capable of binding to their primary ligands, tumor necrosis factor alpha (TNFA) or lymphotoxin alpha (LTA). Upon binding to TNFA, the receptors trimerize, become activated, and transduce intracellular signaling cascades that play a role in various processes, including apoptosis and inflammation. See generally Ward-Kavanagh, LK, et al. (2016). Immunity, 44(5), 1005-1019. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0027] In certain embodiments, any region of the TNFRSF1A gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TNFRSF1A gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TNFRSF1A gene targeted by the RNA-guided nuclease is derived from a human (hTNFRSF1A). In some embodiments, the TNFRSF1A gene targeted by the RNA-guided nuclease is derived from a dog (cTNFRSF1A). In some embodiments, the TNFRSF1A gene targeted by the RNA-guided nuclease is derived from a horse (eTNFRSF1A). In some embodiments, the TNFRSF1A gene targeted by the RNA-guided nuclease is of feline origin (fTNFRSF1A).

[0028] The term "tumor necrosis factor receptor 2" or "TNFRSF1B" refers to a gene (NCBI Gene ID: 7133 [human], NCBI Gene ID: 487437 [dog], NCBI Gene ID: 100055840 [horse], NCBI Gene ID: 101080392 [cat]), or the encoded gene product (e.g., UniProt: P20333; XP_011540362.1 [human], XP_038387905.1 [dog], XP_023491528.1 [horse], XP_023113905.2 [cat]), as well as sequence variants, proteins with conservative amino acid substitutions, and glycoforms thereof. Typically, the proteins encoded by the above-listed genes are transmembrane receptor proteins capable of binding to TNFA or LTA and are involved in pro-survival pathways through downstream activation of the NFkB pathway. See generally Ward-Kavanagh, LK, et al. (2016). Immunity, 44(5), 1005-1019. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0029] In certain embodiments, any region of the TNFRSF1B gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 13, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TNFRSF1B gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TNFRSF1B gene targeted by the RNA-guided nuclease is derived from a human (hTNFRSF1B). In some embodiments, the TNFRSF1B gene targeted by the RNA-guided nuclease is derived from a dog (cTNFRSF1B). In some embodiments, the TNFRSF1B gene targeted by the RNA-guided nuclease is derived from a horse (eTNFRSF1B). In some embodiments, the TNFRSF1B gene targeted by the RNA-guided nuclease is of feline origin (fTNFRSF1B).

[0030] The term "lymphotoxin beta receptor" or "TNFRSF3" refers to a gene (NCBI Gene ID: 4055 [human], NCBI Gene ID: 486728 [dog], NCBI Gene ID: 100059650 [horse], NCBI Gene ID: 101081146 [cat]), or the encoded gene product (e.g., UniProt: P36941; NP_001257916.1 [human], XP_038295148.1 [dog], XP_001492220.3 [horse], XP_003988366.4 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Typically, the proteins encoded by the above-listed genes play roles in signal transduction, lipid metabolism, immune response, and programmed cell death during lymphatic and other organ development. The primary ligands for this receptor include lymphotoxin alpha / beta and tumor necrosis factor ligand superfamily member 14 (TNFSF14). The activity of this receptor has also been associated with carcinogenesis. See generally Seymour, R., et al. (2006). Veterinary Pathology, 43(4), 401-423. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of protein or gene (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0031] In certain embodiments, any region of the TNFRSF3 gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TNFRSF3 gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TNFRSF3 gene targeted by the RNA-guided nuclease is derived from a human (hTNFRSF3). In some embodiments, the TNFRSF3 gene targeted by the RNA-guided nuclease is derived from a dog (cTNFRSF3). In some embodiments, the TNFRSF3 gene targeted by the RNA-guided nuclease is derived from a horse (eTNFRSF3). In some embodiments, the TNFRSF3 gene targeted by the RNA-guided nuclease is of feline origin (fTNFRSF3).

[0032] The terms "OX40 receptor," "OX40," "CD134," or "TNFRSF4" refer to genes (NCBI Gene ID: 7293 [human], NCBI Gene ID: 489600 [dog], NCBI Gene ID: 100066167 [horse], NCBI Gene ID: 493665 [cat]), or encoded gene products (e.g., UniProt: P43489; XP_011540377.1 [human], XP_038520220.1 [dog], XP_001503612.3 [horse], NP_001009200.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Typically, the proteins encoded by the above-listed genes specifically bind to OX40 ligand (OX40L) and are primarily expressed on lymphocytes. The result of this binding is transient downstream intracellular signaling mediated through TRAF2 and other effector proteins that often occurs in response to the presence of antigen. See generally Ward-Kavanagh, LK, et al. (2016). Immunity, 44(5), 1005-1019. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0033] In certain embodiments, any region of the TNFRSF4 gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TNFRSF4 gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TNFRSF4 gene targeted by the RNA-guided nuclease is derived from a human (hTNFRSF4). In some embodiments, the TNFRSF4 gene targeted by the RNA-guided nuclease is derived from a dog (cTNFRSF4). In some embodiments, the TNFRSF4 gene targeted by the RNA-guided nuclease is derived from a horse (eTNFRSF4). In some embodiments, the TNFRSF4 gene targeted by the RNA-guided nuclease is derived from a feline (fTNFRSF4).

[0034] The term "TNF receptor superfamily member 11A" or "TNFRSF11A" refers to the gene (NCBI Gene ID: 8792 [human], NCBI Gene ID: 483957 [dog], NCBI Gene ID: 100056617 [horse], NCBI Gene ID: 101090651 [cat]), or the encoded gene product (e.g., UniProt: Q9Y6Q6; NP_001257878.1 [human], XP_038509502.1 [dog], XP_023503703.1 [horse], XP_023096972.1 [cat]), as well as sequence variants, proteins bearing conservative amino acid substitutions, and glycoforms thereof. Typically, the proteins encoded by the genes listed above are transmembrane proteins that can interact with n various TRAF family proteins, through which the receptor induces activation of the NF-kappaB and MAPK8 / JNK pathways. This receptor, along with its ligand, is a key regulator of interactions between T cells and dendritic cells and an essential mediator of bone metabolism and development. See generally, Xue, JY, et al. (2021). Journal of Bone and Mineral Metabolism, 39(1), 45-53. In some cases, and simply for disambiguation, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to human, canine, equine, and feline forms, respectively).

[0035] In certain embodiments, any region of the TNFRSF11A gene (e.g., the 5' untranslated region [UTR], exon 1, exon 2, exon 3, exon 4, exon 5, exon 6, exon 7, exon 8, exon 9, exon 10, exon 11, exon 12, any intervening intron region, intron / exon junction, 3' UTR, or polyadenylation signal) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, the TNFRSF11A gene targeted by the RNA-guided nuclease is derived from a mammal. In some embodiments, the TNFRSF11A gene targeted by the RNA-guided nuclease is derived from a human (hTNFRSF11A). In some embodiments, the TNFRSF11A gene targeted by the RNA-guided nuclease is derived from a dog (cTNFRSF11A). In some embodiments, the TNFRSF11A gene targeted by the RNA-guided nuclease is of equine origin (eTNFRSF11A). In some embodiments, the TNFRSF11A gene targeted by the RNA-guided nuclease is of feline origin (fTNFRSF11A).

[0036] The term "treatment" refers to obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in terms of completely or partially preventing a disease or its symptoms, and / or therapeutic, in terms of partially or completely curing the disease and / or adverse effects resulting from the disease. For example, a composition, method, or system of the present disclosure may be administered as a prophylactic treatment to a subject with a predisposition to a given condition (e.g., arthritis). As used herein, "treatment" covers any treatment of a disease in a mammal, particularly a human, dog, cat, or horse, and includes (a) preventing the disease from occurring in a subject predisposed to the disease but not yet diagnosed as having the disease, (b) inhibiting the disease, i.e., arresting the onset or progression of the disease, and (c) relieving the disease, i.e., causing regression of the disease and / or alleviating one or more disease symptoms.

[0037] "Treatment" is also meant to encompass the delivery of an agent to provide a pharmacological effect even in the absence of a disease or condition. For example, "treatment" encompasses the delivery of a composition that can induce an immune response or confer immunity in the absence of a disease state, e.g., in the case of a vaccine. It is understood that the compositions and methods of the present disclosure are applicable to treating all mammals, including, but not limited to, human, canine, feline, equine, and bovine subjects.

[0038] The term "therapeutically effective" refers to an amount of a composition or combination of compositions described herein that is sufficient to provide the intended use, including, but not limited to, disease treatment. A therapeutically effective amount may vary depending on the intended use (in vitro or in vivo), or the subject and disease state being treated (e.g., the subject's weight, age, or sex), the severity of the disease state, or the mode of administration. The term also applies to a dose that will induce a specific response in target cells (e.g., reduced platelet adhesion and / or cell migration). A particular dose will vary depending on the particular composition selected, the administration regimen to be followed, whether the composition is administered in combination with other compositions or compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system through which the composition is transported.

[0039] The term "joint disease" is defined as a condition that induces anatomical and / or physiological changes in the joint, e.g., a measurable abnormality in the cells or tissues of the joint that may lead to metabolic and molecular disorders, including, but not limited to, radiological detection of joint space narrowing, subchondral sclerosis, subchondral cysts, and osteophyte formation.

[0040] "Joint disease" is defined in a human subject as a symptom that prompts the subject to seek medical intervention, e.g., pain, stiffness, swelling, or rigidity reported by the subject. For non-human mammals, "joint disease" is defined as, e.g., lameness, observable changes in gait, weight bearing, allodynia, or exploratory behavior.

[0041] "Back or spinal conditions or disorders" include, but are not limited to, lower back pain, neck pain, disc disorders, adolescent idiopathic scoliosis, adult spinal deformity, cervical degenerative disc disease, cervical disc herniation, cervical myelopathy, cervical stenosis, compression fractures, lumbar degenerative spondylolisthesis, lumbar spondylolisthesis, lower back sprains and strains, lumbar degenerative disc disease, lumbar disc herniation, lumbar stenosis, cervical sprains (whiplash) and strains, cervical strains, osteoporosis, and whiplash. Generally, such disorders or conditions contribute to or cause local pain, inflammation, or morphological changes (e.g., fibrosis, degeneration, osteolysis, bone formation) in the cervical, thoracic, lumbar, or sacral spine or surrounding tissues.

[0042] "Lower back pain" is defined as measurable or identifiable pain or discomfort (either chronic or sporadic) in a given subject, encompassing at least the lumbar spinal region of a mammal. The pain may be localized to the lower back (e.g., muscle aches) or may present as shooting, burning, stabbing, and / or radiating pain sensations throughout the subject's back and / or limbs. The pain may be idiopathic or may be associated with one or more underlying conditions (diagnosed or undiagnosed), including, but not limited to, chronic inflammation, arthritis, osteoporosis, trauma (e.g., post-surgery), neuropathy, musculoskeletal abnormalities (e.g., spondylolisthesis or spinal stenosis), herniated nucleus pulposus (HNP), annular ligament rupture, facet arthritis, nerve root compression, and / or degenerative disorders.

[0043] "Neck pain" is defined as measurable or identifiable pain or discomfort associated with the cervical spine or adjacent ligaments, muscles, and / or tendons. The pain may manifest as localized pain in the neck, or as a shooting, stabbing, burning, and / or radiating sensation throughout the back or limbs, including, but not limited to, the subject's head, shoulders, arms, legs, and / or back. Neck pain may be idiopathic or may be associated with one or more underlying conditions (diagnosed or undiagnosed), including, but not limited to, rheumatoid arthritis, osteoporosis, fibromyalgia, chronic inflammation, herniated discs, spondylosis, spinal stenosis, whiplash, and / or degenerative disorders.

[0044] The terms "polynucleotide," "nucleotide," and "nucleic acid" are used interchangeably herein and refer to all forms of nucleic acids, oligonucleotides, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Polynucleotides include genomic DNA, cDNA, and antisense DNA, as well as spliced ​​or unspliced ​​mRNA, rRNA, tRNA, lncRNA, RNA antagonists, and inhibitory DNA or RNA (e.g., RNAi, e.g., small or short hairpin (sh)RNA, microRNA (miRNA), aptamers, small or short interfering (si)RNA, trans-splicing RNA, or antisense RNA). Polynucleotides also include non-coding RNA, including, but not limited to, RNAi, miRNA, lncRNA, RNA antagonists, aptamers, and any other non-coding RNA known to those skilled in the art. Polynucleotides include naturally occurring, synthetic, and intentionally altered or modified polynucleotides, as well as analogs and derivatives. The term "polynucleotide" also refers to a polymeric form of nucleotides of any length, including deoxyribonucleotides or ribonucleotides, or analogs thereof, and is synonymous with nucleic acid sequence. Polynucleotides may contain modified nucleotides, such as methylated nucleotides and nucleotide analogs, and may be interrupted by non-nucleotide components. If present, modifications to the nucleotide structure may be imparted before or after assembly of the polymer. The term polynucleotide, as used herein, refers interchangeably to double- and single-stranded molecules. Unless otherwise specified or required, any embodiment described herein that includes a polynucleotide encompasses both the double-stranded form and two complementary single-stranded forms known or predicted to constitute a double-stranded form. Polynucleotides may be single-, double-, or triplex-stranded, linear or circular, and may be of any length. When discussing polynucleotides, the sequence or structure of a particular polynucleotide may be described herein according to the convention of providing the sequence in the 5' to 3' direction.

[0045] The term "gene" or "nucleotide sequence encoding a polypeptide" refers to a segment of DNA involved in producing a polypeptide chain. The DNA segment can include regions preceding and following the coding region (leader and trailer) involved in the transcription / translation of the gene product and the regulation of transcription / translation, as well as intervening sequences (introns) between individual coding segments (exons). For example, a gene comprises a polynucleotide containing at least one open reading frame that, after being transcribed and translated, is capable of encoding a specific protein or polypeptide.

[0046] The terms "extracellular domain" and "ectodomain" may be used interchangeably and, when referring to a transmembrane cellular receptor, are defined as the portion of a protein that is exposed to the extracellular environment and that can engage and / or bind to a ligand.

[0047] The terms "cytoplasmic domain" and "intracellular domain" may be used interchangeably and, when referring to transmembrane receptors, define the portions of a protein that are exposed to the cytoplasm. Often, these portions of the protein contain signaling domains for recruiting and associating with various intracellular factors. After engagement with a ligand via the extracellular domain, the interaction effect may change, resulting in new association, dissociation, or recruitment of various cytoplasmic factors that help transmit the signal.

[0048] The term "transmembrane domain," which may be abbreviated as "TM" to refer to a transmembrane receptor, is defined as the portion of a protein that is embedded within the plasma membrane (i.e., not exposed to either the extracellular environment or the cytosol). Transmembrane domains generally have more hydrophobic characteristics than either the extracellular or cytoplasmic portions and often adopt a superhelical structure. Although its primary role is as an anchor, ligand-induced conformational changes to certain receptors have been shown to affect the transmembrane domain, making it essential for subsequent intracellular signal transduction.

[0049] The term "receptor" refers to a protein that can bind with high affinity to another cognate protein (i.e., its ligand). This receptor-ligand interaction can be 1:1 or can result in multimerization, where multiple proteins aggregate to bind to one or more ligands. Receptors are generally found on the cell surface so that they can most efficiently encounter ligands and initiate intracellular signaling.

[0050] The term "intracellular signaling" refers to cellular changes resulting from events occurring at the cell surface. Typically, a soluble ligand binds to its receptor on the cell surface, which can induce changes in the receptor, resulting in related intracellular factors being affected. These factors can then affect others within the cell, and this cascade often continues until a specific factor can modify gene expression in the nucleus in response to a surface stimulus.

[0051] The term "RNA-guided nuclease" refers to an enzyme that can, for example, destroy the backbone of a DNA molecule. The activity of an RNA-guided nuclease is guided by a nucleic acid molecule (i.e., a guide RNA). When properly oriented to form a functional ribonucleoprotein complex, the enzyme is localized to a specific location within a target nucleic acid (e.g., a gene or locus) through sequence complementarity with a portion of the guide RNA. Non-exhaustive examples of RNA-guided nucleases include Cas9, Cas12, and Cas12a (formerly known as Cpf1).

[0052] The term "Cas9" refers to an RNA-guided, double-stranded DNA-binding nuclease or nickase protein, or variants thereof, and may be used to refer to either naturally occurring or recombinant Cas9 nuclease variants (e.g., ES-Cas9, HF-Cas9, PE-Cas9, and AR-Cas9). Wild-type Cas9 nucleases have two functional domains, e.g., RuvC and HNH, that simultaneously cleave both strands of double-stranded DNA, resulting in a double-stranded break. The Cas9 enzymes described herein can contain an HNH or HNH-like nuclease domain and / or a RuvC or RuvC-like nuclease domain without affecting their ability to induce a double-stranded break in genomic DNA (e.g., at a target locus) when both functional domains are active. The Cas9 enzyme may comprise one or more catalytic domains of a Cas9 protein from bacteria belonging to the group consisting of Corynebacter, Sutterella, Legionella, Treponema, Filifactor, Eubacterium, Streptococcus, Lactobacillus, Mycoplasma, Bacteroides, Flaviivola, Flavobacterium, Sphaerochaeta, Azospirillum, Gluconacetobacter, Neisseria, Roseburia, Parvibaculum, Staphylococcus, Nitratifractor, and Campylobacter. In some embodiments, the two catalytic domains are from different bacterial species.

[0053] As used herein, "PAM" refers to a protospacer-adjacent motif, which is required for RNA-guided nucleases to bind to target nucleic acids. Often, the PAM is immediately adjacent to the complementary sequence in the target. Native Cas9, e.g., molecules, recognize specific PAM sequences (see, e.g., Table 1). In some embodiments, the Cas9 molecule has the same PAM specificity as a native Cas9 molecule. In other embodiments, the Cas9 molecule has a PAM specificity not associated with a native Cas9 molecule. In other embodiments, the PAM specificity of a Cas9 molecule is not associated with the native Cas9 molecule to which it has closest sequence homology. For example, a native Cas9 molecule can be modified so that PAM sequence recognition is modified to reduce off-target sites, improve specificity, or eliminate the PAM recognition requirement. In one embodiment, the Cas9 molecule can be modified (e.g., to lengthen the PAM recognition sequence, improve Cas9 specificity to a higher level of identity, reduce off-target sites, and / or increase specificity). In one embodiment, the length of the PAM recognition sequence is at least 4, 5, 6, 7, 8, 9, 10, or 15 amino acids in length. In some embodiments, the Cas9 molecule can be engineered to excise PAM recognition.

[0054] The terms "guide RNA," "gRNA," or "sgRNA" may be used interchangeably and refer to an RNA molecule, preferably a synthetic RNA molecule, composed of a targeting (crRNA) sequence and a scaffold. These molecules, when loaded onto a functional RNA-guided nuclease, can direct sequence-specific cleavage of a target nucleic acid.

[0055] The sgRNA can be administered or formulated, for example, as synthetic RNA or as a nucleic acid comprising a sequence encoding the gRNA, which is then expressed in the target cell. As will be apparent to one of skill in the art, various tools can be used in the design and / or optimization of sgRNAs, for example, to increase the specificity and / or precision of genome editing at a particular site.

[0056] Generally, candidate sgRNAs can be designed and identified by first locating a suitable PAM within the genome sequence. Additional calculations can then be used to predict on-target and off-target efficiency. Available web-based tools to aid in the initial setup and modeling of candidate sgRNAs include, but are not limited to, CRISPRseek, CRISPR Design Tool, Cas-OFFinder, E-CRISP, ChopChop, CasOT, CRISPR direct, CRISPOR, BREAKING-CAS, CrispRGold, and CCTop. See, for example, Safari, F. et al. (2017) Current Pharmaceutical Biotechnology, 18(13):1038-54, which is incorporated herein by reference in its entirety for all purposes. Such tools are also described, for example, in PCT Publication No. WO2014 / 093701A1 and Liu, G. et al. (2020). Computational approaches for effective CRISPR guide RNA design and evaluation. Computational and Structural Biotechnology Journal, 18:35-44, each of which is incorporated by reference in its entirety for all purposes. Candidate sgRNAs can be further evaluated by experimental screening or other methodologies.

[0057] The term "CRISPR RNA" or "crRNA" refers to the portion of an sgRNA molecule that has complementarity to a target nucleic acid.

[0058] The phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0059] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and inactive ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Except insofar as any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is incompatible with the active pharmaceutical ingredient, its use in the therapeutic compositions of the present disclosure is contemplated. Additional active pharmaceutical ingredients, such as other drugs, may also be incorporated into the compositions and methods described.

[0060] The term "pharmaceutically acceptable excipient" is intended to include vehicles and carriers that can be co-administered with a compound to facilitate the performance of its intended function. The use of such vehicles for pharmaceutically active substances is well known in the art. Examples of such vehicles and carriers include solutions, solvents, dispersion media, delay agents, emulsions, etc. Any other conventional carriers suitable for use with multibinding compounds also fall within the scope of this disclosure.

[0061] As used herein, the terms "a," "an," or "the" are generally intended to cover both the singular and the plural.

[0062] The terms "about" and "approximately" refer to a statistically significant range of values. Such a range can be within an order of magnitude of a given value or range, preferably within 50%, more preferably within 20%, more preferably still within 10%, and even more preferably within 5%. The acceptable variation encompassed by the terms "about" or "approximately" depends on the particular system under study and can be readily understood by one of ordinary skill in the art. Furthermore, as used herein, the terms "about" and "approximately" mean that compositions, amounts, formulations, parameters, shapes, and other quantities and characteristics are not, and need not be, exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding, measurement error, etc., and other factors known to those of ordinary skill in the art. Generally, a dimension, size, formulation, parameter, shape, or other quantity or characteristic is "about" or "approximately" whether or not explicitly stated as such. Note that embodiments of widely different sizes, shapes, and dimensions may employ the described configurations.

[0063] As used herein, the term "substantially" can refer to a majority or majority, such as at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more.

[0064] When used in the appended claims, in their original and amended forms, the transitional phrases "comprising," "consisting essentially of," and "consisting of" define the claim in terms of which additional unrecited claim elements or steps, if any, are excluded from the claim. The term "comprising" is intended to be inclusive or open-ended and does not exclude any additional unrecited elements, methods, steps, or materials. The term "consisting of" excludes any element, step, or material other than that specified in the claim, and in the latter case, excludes impurities normally associated with the specified material. The term "consisting essentially of" limits the claim to the specified element, step, or material, and to those that do not materially affect the basic and novel characteristics of the claimed methods and compositions. All compositions, methods, and kits described herein that embody the present disclosure may, in alternative embodiments, be more specifically defined by any of the transitional phrases "comprising," "essentially consisting of," and "consisting of."

[0065] III. Method A. CRISPR Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. Additionally, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0066] In one aspect, the present disclosure encompasses compositions related to clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated RNA-guided nucleases, as well as related methods, components, and compositions (hereinafter, CRISPR / Cas systems). Such systems minimally require at least one isolated or non-naturally occurring RNA-guided nuclease (e.g., a Cas9 protein) and at least one isolated or non-naturally occurring guide RNA (e.g., an sgRNA) to effect amplification of a nucleic acid sequence (e.g., genomic DNA).

[0067] In some embodiments, the CRISPR / Cas system effects a sequence modification at a target location (e.g., an insertion or deletion (collectively, indels) that results in a loss-of-function (i.e., knockout) of the affected gene or allele; e.g., a nucleotide substitution that results in a truncation, nonsense mutation, or other loss-of-function of the encoded product of one or more IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, or TNFRSF11A genes (i.e., mRNA or protein); e.g., a truncation, nonsense mutation, or other loss-of-function of the encoded product of one or more IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, or TNFRSF11A genes). loss-of-function of the encoded mRNA or protein, e.g., by causing a single-nucleotide, double-nucleotide, or other frameshift deletion, or a deletion resulting in a premature stop codon; or an insertion that results in a truncation, nonsense mutation, or other loss-of-function of an encoded gene product, such as, for example, the encoded gene product of one or more IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, or TNFRSF11A genes (i.e., mRNA or protein); resulting in the modification of a target gene or locus in a eukaryotic cell. In some embodiments, the CRISPR / Cas systems of the present disclosure provide for the modification of a gene and / or its encoded product such that the modified product has a resulting loss-of-function and is a dominant negative or decoy (e.g., a transmembrane or soluble receptor that is unable to initiate intracellular signaling).

[0068] In one aspect, the CRISPR / Cas system alters the sequence of a nucleic acid through nuclease activity. For example, in the case of genomic DNA, an RNA-guided nuclease is positioned at a target location within a targeted gene or locus by sequence complementarity with the target genomic sequence (e.g., the complementary component of a CRISPR RNA (crRNA) or synthetic single-guide RNA (sgRNA)), and cleaves the genomic DNA upon recognizing a specific nuclease-specific motif called a protospacer adjacent motif (PAM). See generally, Collias, D., & Beisel, CL (2021). Nature Communications, 12(1), 1-12.

[0069] Nuclease activity (i.e., cleavage) induces double-strand breaks (DSBs) in genomic DNA. Endogenous cellular mechanisms for DSB repair, i.e., nonhomologous end joining (NHEJ), microhomology-mediated end joining (MMEJ), and homologous recombination, result in erroneous repair at a given target location with some calculable frequency as a result of interference from the relevant components of the CRISPR / Cas system, thereby introducing substitutions or indels into genomic DNA. See generally Scully, R., et al. (2019). Nature Reviews Molecular Cell Biology, 20(11), 698-714. At some frequency, these indels and / or substitutions can result in frameshifts, nonsense mutations (i.e., premature stop codons), or truncations, which affect the availability of gene products such as mRNA and / or proteins. In certain embodiments, the CRISPR / Cas system can induce a homology-directed repair (HDR) mechanism that leads to the insertion of non-random sequences at the target location through the use of a template (e.g., an HDR template) that is provided to the cell as part of the system along with a nuclease and a gRNA. See Bloh, K., & Rivera-Torres, N. (2021). International Journal of Molecular Sciences, 22(8), 3834.

[0070] Generally, the minimum requirements for a CRISPR / Cas system depend on the nuclease (i.e., Cas protein) provided with it. To this extent, these bacterial nucleases have been functionally divided into types I, III, and V, which are all classified into class 1, and types II, IV, and VI, which are classified into class 2.

[0071] Class 1 CRISPR / Cas systems: The exact components, compositions, and methods for effecting targeted nucleic acid sequence alterations using Class 1 CRISPR / Cas systems vary, but should minimally include a nuclease (selected from at least Type I and Type III), 1) at least one guide RNA selected from sgRNAs, or 2) a combination of crRNA and tracrRNA. These CRISPR / Cas systems are categorized together as Class 1 CRISPR / Cas systems due to their similarities in requirements and modes of action within eukaryotic cells. To this end, compositions, components, and methods among Class 1 components can be considered functionally interchangeable, and the following details, provided solely for illustrative purposes, do not represent an exhaustive list of class members.

[0072] Cas3 (see Table 1) is a prototypical type I DNA nuclease that functions as an effector protein as part of a larger complex (the Cascade complex, which includes Cse1 and Cse2) capable of genome editing. See generally He, L., et al. (2020). Genes, 11(2), 208. Unlike other CRISPR / Cas systems, type I systems localize to DNA targets without the Cas3 nuclease via the Cascade complex, which then recruits Cas3 to cleave DNA upon binding and localizing the 3' PAM. The Cascade complex is also responsible for processing crRNA so that it can be used to guide crRNA to target locations. This functionality gives Cascade the ability to process multiple arrayed crRNAs from a single molecule. See Luo, M. (2015). Nucleic Acids Research, 43(1), 674-681. Thus, type I systems may be used to edit multiple targeted genes or loci from a single molecule.

[0073] Because the natural Cas3 substrate is ssDNA, its function in genome editing is thought to be as a nicases; however, when targeted in tandem, the resulting edits are the result of blunt-end cleavage on opposite strands to approximate blunt-cutting endonucleases such as Cas9. See Pickar-Oliver, A., & Gersbach, CA (2019). Nature Reviews Molecular Cell Biology, 20(8), 490-507.

[0074] Similar to type I nucleases, type III systems rely on a complex of proteins to effect nucleic acid cleavage. In particular, Cas10 possesses nuclease activity for cleaving ssDNA in prokaryotes. See Tamulaitis, G. Trends in Microbiology, 25(1), 49-61 (2017). Interestingly, this archaeal CRISPR / Cas system exhibits dual specificity, targeting both ssDNA and ssRNA. Apart from this change, the system functions very similarly to type I in that the crRNA targets an effector complex (similar to a Cascade) in a sequence-dependent manner. Similarly, the effector complex processes the crRNA before assembly. The dual nature of this nuclease potentially makes its application in genome editing more powerful, as both genomic DNA and, in some cases, mRNA with the same sequence can be targeted to silence specific target genes.

[0075] Class 2 CRISPR / Cas systems: The exact components, compositions, and methods for effecting targeted nucleic acid sequence alterations using Class 2 CRISPR / Cas systems vary, but should minimally include a nuclease (selected from at least Type II and Type V), 1) at least one guide RNA selected from sgRNAs, or 2) a combination of crRNA and tracrRNA. These CRISPR / Cas systems are categorized together as Class 2 CRISPR / Cas systems due to their similarities in requirements and modes of action within eukaryotic cells. To this end, compositions, components, and methods among Class 2 components can be considered functionally interchangeable, and the following details, provided solely for illustrative purposes, do not represent an exhaustive list of class members.

[0076] Type II nucleases are the best-characterized CRISPR / Cas system, particularly the canonical genome-editing nuclease Cas9 (see Table 1). Multiple Cas9 proteins have been isolated from various bacterial species. The main difference between these nucleases is the required recognition site, PAM, within the targeted dsDNA. After association with the gRNA molecule, the crRNA (or the targeting domain of the sgRNA) orients the nuclease to the appropriate location, while protein recognition of the PAM induces a cleavage event near the site, resulting in a blunt DSB.

[0077] In addition to naturally occurring Cas9 proteins, several engineered variants have also been reported. These range from Cas9s with enhanced specificity (i.e., lower off-target activity), such as espCas9. Others have been catalytically modified via point mutations in the RuvC (e.g., D10A) and HNH (e.g., H840A) domains, so that they induce only single-strand breaks (i.e., Cas9 nickase). See Frock, R. et al. (2015). Nature Biotechnology, 33(2), 179-186. They have also been shown to be less error-prone during editing. This reduction in off-target effects is paramount when selecting desired insertions (i.e., knock-in mutations in which the desired nucleotide sequence is introduced into the target nucleic acid molecule) rather than deletions. Indeed, fewer off-target effects may support a preferred DNA repair mechanism (most often HDR for knock-in mutations). See generally Naeem, M., et al. (2020). Cells, 9(7), 1608.

[0078] Additional exemplary further engineered variants (e.g., mutants, chimeras, and the like) of canonical Cas proteins include the following (each of which is incorporated by reference herein in its entirety for all purposes): WO2015 / 035162A2, WO2019 / 126716A1, WO2019 / 126774A1, WO2014 / 093694A1, WO2014 / 150624A1, US2019 / 0225955A1, U.S. Pat. No. 11,427,818, U.S. Pat. No. 11,242,542, U.S. Pat. No. 11,098,297, U.S. Pat. No. 10,876,100, U.S. Pat. No. 10,767,193, U.S. Pat. No. 10,494,621, and U.S. Pat. No. 10,100,291.

[0079] For the avoidance of doubt, SpCas9 collectively refers to any one of the group consisting of espCas9 (also referred to herein as ES-Cas9 or esCas9), HF-Cas9, PE-Cas9, and ARCas9 (also referred to as AR-Cas9).

[0080] Like the canonical Cas9 system, type V nucleases require only a synthetic sgRNA with a targeting domain complementary to the genomic sequence to perform genome editing. These nucleases contain a RuvC domain but lack the HNH domain of type II nucleases. Furthermore, Cas12 leaves a staggered cut in dsDNA substrates distal to the PAM, compared to, for example, the blunt cut of Cas9 next to the PAM. Both Cas12a, also known as Cpf1, and Cas12b, also known as C2c1 (see Table 1), act as part of a larger complex of two gRNA-associated nucleases that act on dsDNA as a quaternary structure, simultaneously nicking each strand. See Zetsche, B. et al. (2015). Cell, 163(3):759-771. See also Liu, L. et al. (2017). Molecular Cell, 65(2):310-322. Furthermore, Cas12b (C2c1) is a highly accurate nuclease with little tolerance for mismatches. See Yang, H. et al. (2016). Cell, 167(7):1814-1828.e12. [Table 1]

[0081] See generally Wang, J., Zhang, C., & Feng, B. (2020). Journal of Cellular and Molecular Medicine, 24(6), 3256-3270, where N = any nucleotide, R = any purine (A or G), Y = any pyrimidine (C or T), W = A or T, and V = A, C, or G.

[0082] In one aspect, the CRISPR / Cas system of the present disclosure comprises at least one RNA-guided nuclease (e.g., Cas protein) from one or more of the following selected bacterial genera: Corynebacterium, Sutterella, Legionella, Treponema, Filifactor, Eubacterium, Streptococcus, Lactobacillus, Mycoplasma, Bacteroides, Flavobacterium, Spirochaeta, Azospirillum, Gluconacetobacter, Neisseria, Roseburia, Parvibaculum, Nitratifractor, Campylobacter, Pseudomonas, Streptomyces, Staphylococcus, Francisella, Acidaminococcus, Lachnospiraceae, Leptotrichia, and Prevotella. In some embodiments, the Cas protein is from a Deltaproteobacteria or Planctomycetes bacterial species.

[0083] Some aspects of the present disclosure provide strategies, methods, compositions, and therapeutic modalities for modifying a targeted sequence within a genetic locus (e.g., modifying the sequence of a wild-type and / or mutant sequence within a cell or mammal) by insertion or deletion of one or more nucleotides mediated by an RNA-guided nuclease and one or more guide RNAs (gRNAs), resulting in loss of function of the targeted gene product. In some embodiments, loss of function results in "knocking out" a gene of interest by eliminating gene expression (i.e., generating a "knockout"). In some embodiments, loss of function results in a non-functional gene product (i.e., a gene product that does not have all the functionality of the wild-type gene product). In some embodiments, loss of function results in expression of a gene product with different properties (e.g., different binding affinity or different cellular localization).

[0084] In certain embodiments, the targeted gene is selected from IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. In some embodiments, any region of the targeted gene (e.g., promoter region, 5' untranslated region, 3' untranslated region, exon, intron, or exon / intron boundary) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, a non-coding region of the targeted gene (e.g., enhancer region, promoter region, intron, 5' UTR, 3' UTR, polyadenylation signal) is targeted to modify the gene.

[0085] CRISPR guide RNA: In one aspect, the CRISPR / Cas system of the present disclosure further provides a gRNA molecule (e.g., an isolated or non-naturally occurring RNA molecule) that interacts with an RNA-guided nuclease. In certain embodiments, the gRNA is an sgRNA that includes a crRNA sequence that includes a nucleotide sequence that is complementary to a sequence in a target nucleic acid. In some embodiments, the sgRNA further includes an RNA scaffold portion (i.e., a tracrRNA) that interacts with the RNA-guided nuclease, such that the crRNA is positioned to scan the target nucleic acid for complementarity. In some embodiments, the system further optionally comprises an oligonucleotide-HDR template with homology on either side of the target position. See Bloh, K., & Rivera-Torres, N. (2021). International Journal of Molecular Sciences, 22(8):3834.

[0086] In one embodiment, the RNA-guided nuclease and sgRNA are configured to orient the associated nuclease such that a cleavage event (e.g., a double-stranded or single-stranded cleavage) occurs in sufficient proximity to a complementary sequence in the targeted nucleic acid, thereby facilitating modification of the nucleic acid sequence. In some embodiments, the crRNA is 20 nucleotides in length. In some embodiments, the crRNA is 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[0087] In some embodiments, the crRNA directs the RNA-guided nuclease so that the cleavage event occurs within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 nucleotides of the complementary sequence in the target nucleic acid. The double-stranded or single-stranded cleavage can be located upstream or downstream of the complementary sequence in the target nucleic acid. In some embodiments, the cleavage event occurs within the targeted gene. In some embodiments, the cleavage event occurs upstream of the targeted gene.

[0088] In certain embodiments, the second gRNA molecule, including the second crRNA, directs the second RNA-guided nuclease so that the cleavage event occurs in sufficient proximity to a complementary sequence in the targeted nucleic acid, thereby facilitating modification of the nucleic acid sequence. In some embodiments, the first gRNA and the second gRNA facilitate a cleavage event within a single targeted gene. In some embodiments, the first gRNA and the second gRNA facilitate cleavage events within different targeted genes. In some embodiments, the second crRNA is 20 nucleotides in length. In some embodiments, the second crRNA is 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[0089] In some embodiments, the second crRNA directs the RNA-guided nuclease so that the cleavage event occurs within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 nucleotides of the complementary sequence in the target nucleic acid. The double-stranded or single-stranded cleavage can be located upstream or downstream of the complementary sequence in the target nucleic acid. In some embodiments, the cleavage event occurs within the targeted gene. In some embodiments, the cleavage event occurs upstream of the targeted gene.

[0090] In some embodiments, the targeting domains of the first gRNA and the second gRNA are configured such that a cleavage event is located within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 nucleotides of the other cleavage event, independently for each gRNA molecule. In some embodiments, the first gRNA and the second gRNA modify the targeting nucleic acid sequence simultaneously. In some embodiments, the first gRNA and the second gRNA modify the targeting nucleic acid sequence sequentially.

[0091] In some embodiments, the single-stranded cleavage is accompanied by a second single-stranded cleavage positioned by the crRNA of the first gRNA and the second gRNA, respectively. For example, the crRNA may direct the associated RNA-guided nuclease so that the cleavage events (e.g., the two single-stranded cleavage events) are positioned within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 nucleotides of each other. In some embodiments, the first crRNA and second crRNA are configured to direct their associated RNA-guided nucleases such that two single-strand breaks occur on opposite strands of genomic DNA, e.g., at the same position or within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50 nucleotides of each other, thereby essentially approximating a double-strand break.

[0092] In some embodiments, the nucleic acids encoding one or more crRNAs are selected from any of SEQ ID NOs: 680-1039. In some embodiments, the nucleic acids encoding one or more crRNAs target hIL1R1 and are selected from any of SEQ ID NOs: 680-824. In some embodiments, the nucleic acids encoding one or more crRNAs target cIL1R1 and are selected from any of SEQ ID NOs: 825-892 and 3336-3420. In some embodiments, the nucleic acids encoding one or more crRNAs target eIL1R1 and are selected from any of SEQ ID NOs: 893-967. In some embodiments, the nucleic acids encoding one or more crRNAs target fIL1R1 and are selected from any of SEQ ID NOs: 968-1039.

[0093] In some embodiments, the nucleic acids encoding one or more crRNAs are selected from any of SEQ ID NOs: 1040-1424. In some embodiments, the nucleic acids encoding one or more crRNAs target hIL1RAP and are selected from any of SEQ ID NOs: 1040-1203. In some embodiments, the nucleic acids encoding one or more crRNAs target cIL1RAP and are selected from any of SEQ ID NOs: 1204-1271 and 3421-3490. In some embodiments, the nucleic acids encoding one or more crRNAs target eIL1RAP and are selected from any of SEQ ID NOs: 1272-1348. In some embodiments, the nucleic acids encoding one or more crRNAs target fIL1RAP and are selected from any of SEQ ID NOs: 1349-1424.

[0094] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 1425-1546. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTGFBR1 and is selected from any of SEQ ID NOs: 1425-1546.

[0095] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 1547-1745. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTGFBR2 and is selected from any of SEQ ID NOs: 1547-1745.

[0096] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 1746-1968. In some embodiments, the nucleic acid encoding one or more crRNAs targets hIL6R and is selected from any of SEQ ID NOs: 1746-1968.

[0097] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 1969-2178. In some embodiments, the nucleic acid encoding one or more crRNAs targets hIL6ST and is selected from any of SEQ ID NOs: 1969-2178.

[0098] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 2179-2395. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTNFRSF1A and is selected from any of SEQ ID NOs: 2179-2395.

[0099] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 2396-2642. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTNFRSF1B and is selected from any of SEQ ID NOs: 2396-2642.

[0100] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 2643-2866. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTNFRSF3 and is selected from any of SEQ ID NOs: 2643-2866.

[0101] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 2867-3041. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTNFRSF4 and is selected from any of SEQ ID NOs: 2867-3041.

[0102] In some embodiments, the nucleic acid encoding one or more crRNAs is selected from any of SEQ ID NOs: 3042-3335. In some embodiments, the nucleic acid encoding one or more crRNAs targets hTNFRSF11A and is selected from any of SEQ ID NOs: 3042-3335.

[0103] In some embodiments, the nucleic acid encodes a second sgRNA molecule. In some embodiments, the nucleic acid encodes a third sgRNA molecule. In some embodiments, the nucleic acid encodes a fourth sgRNA molecule.

[0104] In certain embodiments, the nucleic acid may include (a) a sequence encoding a first sgRNA molecule comprising a crRNA complementary to a sequence in a targeting gene, (b) a sequence encoding a second sgRNA comprising a crRNA complementary to a sequence in a second targeting gene, and (c) a sequence encoding an RNA-guided nuclease (e.g., Cas9). Optionally, (d) and (e) are sequences encoding a third sgRNA and a fourth sgRNA, respectively. In some embodiments, the second targeting gene is the same as the first targeting gene. In other embodiments, the second targeting gene is different from the first targeting gene. In some embodiments, (a), (b), and (c) are encoded within the same nucleic acid molecule (e.g., the same vector). In some embodiments, (a) and (b) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), and (d) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), and (e) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), (d), and (e) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), and (c) are encoded within separate nucleic acid molecules. When three or more sgRNAs are used, any combination of (a), (b), (c), (d), and (e) can be encoded within a single or separate nucleic acid molecule.

[0105] In one aspect, a nucleic acid molecule (i.e., encoding (a), (b), (c), (d), or (e)) is delivered to a target cell (i.e., any combination of the encoded RNA-guided nuclease of (c) and at least one encoded gRNA molecule of (a), (b), (d), or (e) contacts the target cell). In some embodiments, the nucleic acid molecule is delivered to the target cell in vivo. In other embodiments, the nucleic acid molecule is delivered to the target cell ex vivo. In some embodiments, the nucleic acid molecule is delivered to the target cell in vitro. In certain embodiments, the nucleic acid molecule is delivered to the target cell as DNA. In other embodiments, the nucleic acid molecule is delivered to the target cell as RNA (e.g., mRNA). In some embodiments, the product of the nucleic acid molecule is delivered as an assembled ribonucleoprotein (RNP).

[0106] In some embodiments, contacting the target cell comprises delivering the RNA-guided nuclease of (c) as a protein along with at least one nucleic acid molecule of interest selected from (a), (b), (d), and (e). In some embodiments, contacting the target cell comprises delivering the encoded RNA-guided nuclease of (c) as DNA along with at least one nucleic acid molecule of interest selected from (a), (b), (d), and (e). In some embodiments, contacting the target cell comprises delivering the encoded RNA-guided nuclease of (c) as mRNA along with at least one nucleic acid molecule of interest selected from (a), (b), (d), and (e).

[0107] In certain embodiments, CRISPR components are delivered to target cells via nanoparticles. Exemplary nanoparticles that may be used with any of the CRISPR / Cas systems disclosed herein include at least lipid nanoparticles or liposomes, hydrogel nanoparticles, metal-organic nanoparticles, gold nanoparticles, magnetic nanoparticles, and virus-like particles. See generally, Xu, CF et al. (2021). Advanced Drug Delivery Reviews, 168:3-29.

[0108] B.TALEN In one aspect, the present disclosure contemplates the use of methods, components, and compositions relating to nucleic acid sequences (e.g., transcription activator-like effector nucleases (TALENs)) to effect enhancement of targeted genes.

[0109] TALE stands for "Transcription Activator-Like Effector" protein, including TALENs ("Transcription Activator-Like Effector Nucleases"). Methods using the TALE system for gene editing may also be referred to herein as TALE methods. TALEs are naturally occurring proteins derived from plant pathogenic bacteria of the genus Xanthomonas. They contain a DNA-binding domain composed of a series of 33-35 amino acid repeat domains, each of which recognizes a single base pair. TALE specificity is determined by two hypervariable amino acids known as repeat variable dinucleotides (RVDs). Modular TALE repeats are linked together to recognize consecutive DNA sequences. Specific RVDs within the DNA-binding domain recognize bases within the target locus, providing the structural features that assemble a predictable DNA-binding domain. The DNA-binding domain of a TALE is fused to the catalytic domain of a type IIS FokI endonuclease to create a targetable TALE nuclease. To induce site-specific mutagenesis, two individual TALEN arms, separated by a 14-20 base pair spacer region, bring the FokI monomers into close proximity, allowing them to dimerize and generate the target double-strand break.

[0110] Several large-scale systematic studies utilizing various assembly methods have shown that TALE repeats can be combined to recognize virtually any user-defined sequence. Custom-designed TALE arrays are also commercially available through Cellectis Bioresearch (Paris, France), Transposagen Biopharmaceuticals (Lexington, KY, USA), and Life Technologies (Grand Island, NY, USA). TALE and TALEN methods suitable for use in the present disclosure are described in U.S. Patent Application Publication Nos. 2011 / 0201118A1, 2013 / 0117869A1, 2013 / 0315884A1, 2015 / 0203871A1, and 2016 / 0120906A1, the disclosures of which are incorporated herein by reference.

[0111] Non-limiting examples of genes that can be silenced or inhibited by permanent gene editing via the TALE method include IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. Non-limiting examples of genes that can be enhanced by permanent gene editing via the TALE method so that their resulting product functions as a decoy or dominant negative include IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. Non-limiting examples of genes that can be enhanced by permanent gene editing via the TALE method include IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. In one aspect, the present disclosure provides compositions for upregulation of protein receptors (including wild-type or gene-edited), including those that bind anti-inflammatory cytokines via the TALE method.

[0112] Examples of systems, methods, and compositions for modifying expression of target gene sequences by the TALE method and that may be used in accordance with embodiments of the present disclosure are described in U.S. Patent No. 8,586,526, which is incorporated herein by reference.

[0113] C. Zinc Finger Nucleases (ZFNs) In one aspect, the present disclosure contemplates the use of zinc finger nuclease (ZFN)-related methods, components, and compositions to effect the amplification of nucleic acid sequences (e.g., targeting genes).

[0114] Each zinc finger contains approximately 30 amino acids in a conserved ββα configuration. Several amino acids on the surface of the α-helix typically contact 3 bp of the major groove of DNA with varying levels of selectivity. Zinc fingers have two protein domains. The first domain is a DNA-binding domain that contains zinc fingers and is found in eukaryotic transcription factors. The second domain is a nuclease domain that contains the FokI restriction enzyme and is responsible for catalytic cleavage of DNA.

[0115] The DNA-binding domain of an individual ZFN typically contains three to six individual zinc finger repeats, each capable of recognizing 9 to 18 base pairs. If the zinc finger domains are specific for their intended target sites, a pair of three-finger ZFNs recognizing a total of 18 base pairs could theoretically target a single locus in a mammalian genome. One method for generating new zinc finger arrays is to combine smaller zinc finger "modules" of known specificity. The most common modular assembly process involves combining three distinct zinc fingers, each capable of recognizing a 3-base pair DNA sequence, to generate a three-finger array capable of recognizing a 9-base pair target site. Alternatively, selection-based approaches such as oligomerization pool engineering (OPEN) can be used to select new zinc finger arrays from randomized libraries that consider context-dependent interactions between adjacent fingers. Engineered zinc fingers are commercially available, and Sangamo Biosciences (Richmond, CA, USA) has partnered with Sigma-Aldrich (St. Louis, MO, USA) to develop a unique platform (CompoZr®) for zinc finger construction.

[0116] Non-limiting examples of genes that can be silenced or inhibited by permanent gene editing via zinc finger technology include IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. Non-limiting examples of genes that can be enhanced by permanent gene editing via zinc finger technology so that their resulting products function as decoys or dominant negatives include IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. Non-limiting examples of genes that can be enhanced by permanent gene editing via zinc finger technology include IL1R1, IL1RAP, TGFBR1, TGFBR2, IL6R, IL6ST, TNFRSF1A, TNFRSF1B, TNFRSF3, TNFRSF4, TNFRSF11A, and combinations thereof. In one aspect, the present disclosure provides compositions for upregulation of protein receptors (including wild-type or gene-edited), including those that bind anti-inflammatory cytokines via zinc finger technology.

[0117] Examples of systems, methods, and compositions for modifying expression of target gene sequences by zinc finger technology and that may be used in accordance with embodiments of the present disclosure are described in U.S. Patent Nos. 6,534,261, 6,607,882, 6,746,838, 6,794,136, 6,824,978, 6,866,997, 6,933,113, Nos. 6,979,539, 7,013,219, 7,030,215, 7,220,719, 7,241,573, 7,241,574, 7,585,849, 7,595,376, 6,903,185, and 6,479,626, which are incorporated herein by reference.

[0118] Other examples of systems, methods, and compositions for modifying expression of target gene sequences by zinc finger technology and that may be used in accordance with embodiments of the present disclosure are described in Beane, et al., Mol. Therapy, 2015, 23 1380-1390, the disclosure of which is incorporated herein by reference.

[0119] IV. Joint disorders or illnesses A. Introduction As described herein, embodiments of the present disclosure provide compositions and methods for improving joint function and treating joint diseases. In certain embodiments, compositions and methods are provided for gene editing synovial fibroblasts, synoviocytes, chondrocytes, tissue (resident) macrophages, or other cells to reduce pro-inflammatory signaling mediated by the binding of inflammatory cytokines, including, but not limited to, IL1α, IL1β, TNFα, IL6, IL8, IL18, IL33, matrix metalloproteinases (MMPs), TGFβ1, TGFβ2, and combinations thereof, to their cognate receptors. Some embodiments are used to treat various forms of arthritis and other inflammatory joint diseases. Some embodiments are further useful for treating canine lameness due to osteoarthritis. Some embodiments are further useful for treating equine lameness due to joint disease. Some embodiments are further useful for treating feline lameness due to joint disease. Some embodiments are also useful for treating post-traumatic arthritis, gout, pseudogout, psoriatic arthritis, and other inflammatory- or immune-mediated joint diseases.

[0120] Treatment of osteoarthritis, degenerative joint disease, and other joint dysfunction is complex, with few long-term options for either symptom relief or restoration of joint function. Osteoarthritis (OA) is a leading cause of disability due to pain. See Neogi, T. (2013). Osteoarthritis Cartilage, 21(9):1145-53. OA and similar diseases affect all mammalian species, including working animals, domestic pets, and their owners. A common mechanistic feature in joint diseases is the acute presence of chronic inflammation driven by elevated levels of pro-inflammatory cytokine signaling. Depending on the extent of disease progression, joint diseases tend to follow a progressive course that encompasses discomfort, pain, and, in the case of OA in particular, disability.

[0121] Psoriatic arthritis (PsA) is another chronic inflammatory joint disease in which joint symptoms are accompanied by skin lesions, such as those commonly associated with psoriasis. See Boehncke, W. et al. (2014). British Journal of Dermatology, 170(4):772-786. Like other forms of arthritis, such as OA, PsA is driven by pro-inflammatory signaling of a host of cytokines, including IL1. Indeed, PsA susceptibility has been shown to correlate with single nucleotide polymorphisms (SNPs) affecting the activity of the IL1 gene locus. See Rahman, P. et al. (2006). Arthritis and Rheumatism, 54(7):2321-2325. These studies also implicate inflammatory cytokine signaling, generally, and IL1 more specifically, in disease progression.

[0122] Gout is a chronic inflammatory condition affecting the joints. The underlying cause is monosodium urate (MSU) crystal deposition, particularly in articular structures (as well as subcutaneous tissue and other sites), and the resulting host response. See Dalbeth, N., & Stamp, L. (2014). Annals of the Rheumatic Diseases, 73(9):1598-1600. Clinical symptoms include recurrent acute flares of severe inflammatory arthritis and tenosynovitis. IL1 and other pro-inflammatory mediators are major contributors to this host response. See Dinarello, CA (2014). Molecular Medicine, 20(1):S43-S58. To this end, effective blockade of these signaling pathways may provide relief to gout patients.

[0123] Current standard treatments for many patients with joint diseases include anti-inflammatory drugs (e.g., NSAIDs) or anti-rheumatic drugs (e.g., methotrexate [an inhibitor of AICAR] or adalimumab [an anti-TNF-alpha monoclonal antibody]). See Friedman, B., & Cronstein, B. (2019). Joint Bone Spine, 86(3):301-307. All of these treatments require repeated administration for continued effectiveness, which can lead to toxicity issues or resistance over time. Therefore, there is a great need for new methods and compositions for treating joint diseases and conditions to treat these chronic conditions.

[0124] In one aspect, the compositions and methods described herein are directed to treating a joint disease or condition in a mammal in need thereof. In some embodiments, the joint disease or condition is osteoarthritis. In some embodiments, the joint disease or condition is psoriatic arthritis. In some embodiments, the joint disease or condition is gout.

[0125] Advantages of the present disclosure over currently available treatments for mammals suffering from one or more joint diseases or conditions include a longer period of symptomatic relief. Upon gene editing of cells within a joint, pro-inflammatory signaling is silenced through targeted genes for the lifespan of the cell and any mitotic progeny. In contrast, biological treatments require regular administration, which can intensify the impact of a host of potentially severe side effects. Among various genetic approaches, the present disclosure is also superior due to, among other reasons, its resistance to leakiness by modifying protein receptors rather than eliminating ligand expression, which can result in compensatory effects (e.g., the accumulation of other factors due to a lack of negative feedback).

[0126] In some embodiments, the disclosure includes a method for treating or preventing a joint disease or condition in a subject in need thereof, the method comprising administering to a joint of the subject a therapeutically effective amount of a pharmaceutical composition comprising a clustered regularly interspaced short palindromic repeats (CRISPR) gene editing system, the system comprising: (i) a CRISPR-associated (Cas) protein; and (ii) at least one guide RNA targeting the IL1R1 gene, the IL1RAP gene, the TGFBR1 gene, the TGFBR2 gene, the IL6R gene, the IL6ST gene, the TNFRSF1A gene, the TNFRSF1B gene, the TNFRSF3 gene, the TNFRSF4 gene, or the TNFRSF11A gene, or a combination thereof. In some embodiments, the joint disease or condition is osteoarthritis. In some embodiments, the joint disease or condition is psoriatic arthritis. In some embodiments, the joint disease or condition is gout.

[0127] In some embodiments, the present disclosure includes a method for treating or preventing arthritis. Non-limiting examples of arthritis that can be treated using the compositions and methods described herein include post-traumatic arthritis, osteoarthritis (a degenerative condition that affects joints, most commonly the hips, knees, and hands), rheumatoid arthritis (an autoimmune disorder that causes inflammation in joints and surrounding tissues), psoriatic arthritis (a type of arthritis that occurs in people with psoriasis, a skin condition characterized by scaly red patches), gout (a type of arthritis caused by the accumulation of uric acid crystals in the joints), lupus (a chronic autoimmune disorder that can cause inflammation and damage to joints and other organs), ankylosing spondylitis (a type of arthritis that primarily affects the spine and causes inflammation and stiffness), reactive arthritis (a type of arthritis that occurs as a reaction to an infection in the body), septic arthritis (a type of arthritis caused by an infection in the joints), juvenile idiopathic arthritis (a form of arthritis that affects children under 16 years of age), and fibromyalgia (a chronic pain disorder that can cause widespread pain and stiffness, including within the joints).

[0128] In some embodiments, the present disclosure includes methods for the treatment or prevention of pseudogout, crystal arthropathy (e.g., caused by the formation of crystals in joints, such as gout and pseudogout), or CPPD disease (calcium pyrophosphate deposition disease), also known as chondrocalcinosis.

[0129] In some embodiments, the disclosure includes methods for the treatment or prevention of rheumatoid arthritis, psoriasis, asthma, inflammatory bowel disease, multiple sclerosis, Alzheimer's disease, type 2 diabetes, cardiovascular disease, or cancer. In some embodiments, these disorders are treated by administering a CRISPR composition described herein that targets an IL1 receptor, e.g., IL1R1 or IL1AP.

[0130] B. Osteoarthritis In one aspect, the present disclosure encompasses a treatment for osteoarthritis (OA). In some embodiments, the OA treatment comprises a therapeutically effective amount of a clustered regularly interspaced short palindromic repeats (CRISPR) gene editing system, the system comprising (i) a CRISPR-associated (Cas) protein and (ii) at least one guide RNA targeting IL1R1. In some embodiments, the OA treatment comprises a CRISPR gene editing system targeting hIL1R1. In some embodiments, the OA treatment comprises a CRISPR gene editing system targeting cIL1R1. In some embodiments, the OA treatment comprises a CRISPR gene editing system targeting eIL1R1. In some embodiments, the OA treatment comprises a CRISPR gene editing system targeting fIL1R1.

[0131] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of hIL1R1.In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 19 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 20 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 21 of hIL1R1.

[0132] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of cIL1R1.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of cIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of cIL1R1.

[0133] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of eIL1R1.In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of eIL1R1.

[0134] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of fIL1R1.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of fIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of fIL1R1.

[0135] In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting IL1RAP. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hIL1RAP. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cIL1RAP. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eIL1RAP. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fIL1RAP.

[0136] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of hIL1RAP.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of hIL1RAP.

[0137] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of cIL1RAP.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of cIL1RAP.

[0138] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of eIL1RAP.

[0139] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of fIL1RAP.

[0140] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTGFBR1.

[0141] In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of hTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of hTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of hTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of hTGFBR1.

[0142] In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTGFBR1.

[0143] In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of eTGFBR1.

[0144] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fTGFBR1.

[0145] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTGFBR2.

[0146] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of hTGFBR2.

[0147] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTGFBR2.

[0148] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTGFBR2.

[0149] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTGFBR2.

[0150] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting IL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fIL6R.

[0151] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of hIL6R.In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of hIL6R.

[0152] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of cIL6R.

[0153] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of eIL6R.

[0154] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fIL6R.

[0155] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting IL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fIL6ST.

[0156] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of hIL6ST.In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 19 of hIL6ST.

[0157] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of cIL6ST.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of cIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of cIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of cIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of cIL6ST.

[0158] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of eIL6ST.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of eIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of eIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of eIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of eIL6ST.

[0159] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 12 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 13 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 14 of fIL6ST.In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 19 of fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 20 of fIL6ST.

[0160] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF1A.

[0161] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF1A.

[0162] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF1A.

[0163] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF1A.

[0164] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF1A.

[0165] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF1B.

[0166] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hTNFRSF1B.

[0167] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF1B.

[0168] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of eTNFRSF1B.

[0169] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF1B.

[0170] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF3.

[0171] In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hTNFRSF3.

[0172] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF3.

[0173] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF3.

[0174] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF3.

[0175] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF4.

[0176] In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hTNFRSF4. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hTNFRSF4. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hTNFRSF4. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hTNFRSF4. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hTNFRSF4. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hTNFRSF4.

[0177] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF4.

[0178] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of eTNFRSF4.

[0179] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF4.

[0180] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF11A.

[0181] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hTNFRSF11A.

[0182] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF11A. In some embodiments, a CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF11A.

[0183] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of eTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF11A.

[0184] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fTNFRSF11A.

[0185] C. Psoriatic arthritis In one aspect, the present disclosure encompasses a treatment for psoriatic arthritis (PsA). In some embodiments, the psoriatic arthritis treatment comprises a therapeutically effective amount of a clustered regularly interspaced short palindromic repeats (CRISPR) gene editing system, the system comprising (i) a CRISPR-associated (Cas) protein and (ii) at least one guide RNA targeting IL1R1. In some embodiments, the psoriatic arthritis treatment comprises a CRISPR gene editing system targeting hIL1R1. In some embodiments, the psoriatic arthritis treatment comprises a CRISPR gene editing system targeting cIL1R1. In some embodiments, the psoriatic arthritis treatment comprises a CRISPR gene editing system targeting eIL1R1. In some embodiments, the psoriatic arthritis treatment comprises a CRISPR gene editing system targeting fIL1R1.

[0186] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of hIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of hIL1R1.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 19 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 20 of hIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 21 of hIL1R1.

[0187] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of cIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of cIL1R1.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of cIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of cIL1R1.

[0188] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of eIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of eIL1R1.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of eIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of eIL1R1.

[0189] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of fIL1R1. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of fIL1R1.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of fIL1R1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of fIL1R1.

[0190] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting IL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting fIL1RAP.

[0191] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of hIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of hIL1RAP.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of hIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of hIL1RAP.

[0192] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of cIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of cIL1RAP.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of cIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of cIL1RAP.

[0193] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of eIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of eIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of eIL1RAP.

[0194] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of fIL1RAP. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of fIL1RAP. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of fIL1RAP.In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of fIL1RAP.

[0195] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting TGFBR1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting hTGFBR1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting cTGFBR1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting eTGFBR1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting fTGFBR1.

[0196] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hTGFBR1.

[0197] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cTGFBR1.

[0198] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of eTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of eTGFBR1.

[0199] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of fTGFBR1. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of fTGFBR1.

[0200] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting TGFBR2. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting hTGFBR2. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting cTGFBR2. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting eTGFBR2. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting fTGFBR2.

[0201] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hTGFBR2.

[0202] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cTGFBR2.

[0203] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eTGFBR2.

[0204] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fTGFBR2. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fTGFBR2.

[0205] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting IL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting fIL6R.

[0206] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hIL6R. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hIL6R. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hIL6R. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hIL6R. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hIL6R. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hIL6R. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of hIL6R.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of hIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of hIL6R.

[0207] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of cIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of cIL6R.

[0208] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of eIL6R.

[0209] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of fIL6R. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of fIL6R.

[0210] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting IL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting fIL6ST.

[0211] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of hIL6ST.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of hIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 19 of hIL6ST.

[0212] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of cIL6ST.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of cIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of cIL6ST.

[0213] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of eIL6ST.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of eIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of eIL6ST.

[0214] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of fIL6ST.In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of fIL6ST. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 19 of fIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 20 of fIL6ST.

[0215] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting TNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting fTNFRSF1A.

[0216] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hTNFRSF1A.

[0217] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of cTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of cTNFRSF1A.

[0218] In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eTNFRSF1A. In some embodiments, a CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs ...

Claims

1. 1. A pharmaceutical composition for treating a disorder having symptoms caused at least in part by intercellular signaling mediated through a transmembrane receptor, comprising: (i) an RNA-guided nuclease or a nucleic acid encoding an RNA-guided nuclease; (ii) at least one guide RNA or a nucleic acid encoding at least one guide RNA that targets the gene encoding the transmembrane receptor.

2. The pharmaceutical composition of claim 1 , wherein the disorder is a musculoskeletal disorder.

3. 2. The pharmaceutical composition of claim 1, wherein the transmembrane receptor is an interleukin-1 receptor selected from the group consisting of IL1R1, IL1RAP, IL1R5, IL1R6, IL1R7, and IL1R9.

4. 4. The pharmaceutical composition of claim 3, wherein the at least one guide RNA targets the IL1RAP gene.

5. 5. The pharmaceutical composition of claim 4, wherein the at least one guide RNA comprises a crRNA sequence selected from the group consisting of SEQ ID NOs: 1040-1194, 1204-1271, 3421-3490, 1272-1348, 1349-1424, and 3514-3543.

6. 2. The pharmaceutical composition of claim 1, wherein the transmembrane receptor is an interleukin-6 receptor.

7. 7. The pharmaceutical composition of claim 6, wherein the at least one guide RNA targets the IL6ST gene.

8. 8. The pharmaceutical composition of claim 7, wherein the at least one guide RNA comprises a crRNA sequence selected from the group consisting of SEQ ID NOs: 1969-2176.

9. 8. The pharmaceutical composition of claim 7, wherein the at least one guide RNA comprises a crRNA sequence selected from the group consisting of SEQ ID NOs: 3567-3606.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the RNA-guided nuclease or the nucleic acid encoding the RNA-guided nuclease is an mRNA encoding the RNA-guided nuclease.

11. The pharmaceutical composition of any one of claims 1 to 9, wherein the RNA-guided nuclease is a Cas protein.

12. 12. The pharmaceutical composition of claim 11, wherein the Cas protein is a Cas9 protein.

13. 12. The pharmaceutical composition of claim 11, wherein the Cas9 protein is a S. pyogenes Cas9 polypeptide.

14. The pharmaceutical composition according to any one of claims 1 to 9, wherein the at least one guide RNA or the nucleic acid encoding at least one guide RNA is the at least one guide RNA.

15. The pharmaceutical composition of any one of claims 1 to 9, wherein the at least one guide RNA is a single guide RNA (sgRNA).

16. The pharmaceutical composition of any one of claims 1 to 9, wherein the at least one guide RNA targets a human gene.

17. The pharmaceutical composition of any one of claims 1 to 9, wherein the at least one guide RNA targets a canine gene.

18. 10. The pharmaceutical composition of claim 1, wherein the composition comprises one or more lipid nanoparticles (LNPs) collectively comprising (i) an RNA-guided nuclease or a nucleic acid encoding an RNA-guided nuclease, and (ii) at least one guide RNA or a nucleic acid encoding at least one guide RNA that targets a gene encoding the transmembrane receptor.

19. 1. Use of a composition in the manufacture of a medicament for treating a disorder in a subject in need thereof, the disorder having symptoms caused at least in part by intracellular signaling mediated through a transmembrane receptor, comprising: The composition comprises: (i) an RNA-guided nuclease or a nucleic acid encoding an RNA-guided nuclease; (ii) at least one guide RNA or a nucleic acid encoding at least one guide RNA targeting the gene encoding the transmembrane receptor; The above use.

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