Lipid nanoparticles for gene editing systems

Abstract

The present disclosure provides compositions and methods for treating and preventing localized pain, inflammation, or morphological changes associated with joint diseases or disorders, back or spinal conditions or disorders, and musculoskeletal diseases or dysfunctions with LNP-encapsulated CRISPR / Cas9 gene editing systems.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 334,476, filed April 25, 2022, U.S. Provisional Patent Application No. 63 / 362,858, filed April 12, 2022, U.S. Provisional Patent Application No. 63 / 342,471, filed May 16, 2022, and U.S. Provisional Patent Application No. 63 / 495,461, filed April 11, 2023, the contents of which are incorporated herein by reference in their entirety for all purposes. [Background technology]

[0002] joint disorders Treatment of osteoarthritis, degenerative joint disease, and other joint dysfunction is complex, and few long-term options exist for symptomatic relief or restoration of joint function. Osteoarthritis (OA) is a leading cause of pain-related disability. Neogi, Osteoarthritis Cartilage 2013;21:1145-53. All mammalian species are affected: working animals, livestock, and their owners all suffer from OA-related discomfort, pain, and disability, depending on the extent of disease progression.

[0003] OA is a complex disease characterized by a progressive and disabling course. Systemic inflammation is associated with OA and OA disease progression. Inflammation is caused by increased levels of pro-inflammatory cytokines. New methods and compositions for treating this disease are urgently needed. Disclosed herein are compositions and methods useful for treating OA and other inflammatory joint disorders.

[0004] Back and spinal disorders Back or spinal conditions or disorders, including lower back pain and pain or inflammation associated with discogenic disorders such as degenerative disc disease (DDD) or internal disc disruption (IDD), are a major cause of morbidity and disability worldwide, and few long-term options currently exist for their improvement. Andersson GB. Epidemiological features of chronic low-back pain. Lancet. 1999;354:581-585. Currently available treatments include surgical or minimally invasive options that often fail to provide long-term relief. Ju, et al. Global Spine Journal (2020):2192568220963058. All vertebrate species are affected by back or spinal conditions or disorders, including working animals, household pets, and their owners. All suffer from associated discomfort, pain, and disability, depending on the degree of disease progression.

[0005] Back or spinal conditions or disorders, such as lower back pain, are complex diseases characterized by numerous factors that contribute to a progressively disabling course. These contributing factors include morphological irregularities (e.g., disc disruption), inflammation, and changes in the local cellular environment (e.g., angiogenesis and / or innervation). Peng, Bao-Gan. World Journal of Orthopedics 4.2 (2013): 42. Each contributing factor is driven by the differential expression of various gene products, including at least pro-inflammatory cytokines, growth factors, and other effector biomolecules. New methods and compositions for treating this disease are urgently needed. Summary of the Invention

[0006] Compositions and methods for treating synovial joint dysfunction are provided and described herein. Additionally, compositions and methods for treating or preventing local pain, inflammation, or morphological changes associated with back or spinal conditions or disorders are disclosed herein. Further, compositions and methods for treating or preventing musculoskeletal diseases and dysfunctions, including, for example, fibrosis and / or scarring in post-operative subjects, are described herein. Additionally, methods for gene editing of cells, including, but not limited to, synovial cells and / or synovial cells, chondrocytes, synovial macrophages, and synovial fibroblasts, and the use of gene-edited synovial cells and / or synovial cells, chondrocytes, synovial macrophages, and synovial fibroblasts in the treatment of diseases such as osteoarthritis are disclosed herein. [Brief explanation of the drawings]

[0007] 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.

[0008] [Figure 1] SEQ ID NOS: 1-48 are crRNA sequences generated by the bioinformatic methods described herein that target human ADAM17 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 2] SEQ ID NOS: 49-96 are crRNA sequences generated by the bioinformatic methods described herein that target human ADAMTS1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and a score, summarizing several predicted performance indicators. [Figure 3]SEQ ID NOS: 97-144 are crRNA sequences generated by the bioinformatic methods described herein that target human ADAMTS5 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 4] SEQ ID NOS:145-192 are crRNA sequences generated by the bioinformatic methods described herein that target human ADM to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 5] SEQ ID NOS:193-240 are crRNA sequences generated by the bioinformatic methods described herein that target human ATP1A1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 6] SEQ ID NOS:241-281 are crRNA sequences generated by the bioinformatic methods described herein that target human BDNF to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 7] SEQ ID NOS:282-301 are crRNA sequences generated by the bioinformatic methods described herein that target human CALCA to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 8]SEQ ID NOS:302-318 are crRNA sequences generated by the bioinformatic methods described herein that target human CALCB to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 9] SEQ ID NOS:319-340 are crRNA sequences generated by the bioinformatic methods described herein that target human CALCRL to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 10] SEQ ID NOS:341-357 are crRNA sequences generated by the bioinformatic methods described herein that target human CCL2 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 11] SEQ ID NOS:358-374 are crRNA sequences generated by the bioinformatic methods described herein that target human CCL3 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 12] SEQ ID NOS:375-391 are crRNA sequences generated by the bioinformatic methods described herein that target human CCL5 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 13]SEQ ID NOS:392-408 are crRNA sequences generated by the bioinformatic methods described herein that target human CCL7 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 14] SEQ ID NOS:409-425 are crRNA sequences generated by the bioinformatic methods described herein that target human CCL20 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 15] SEQ ID NOS:426-473 are crRNA sequences generated by the bioinformatic methods described herein that target human CCN2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 16] SEQ ID NOS:474-517 are crRNA sequences generated by the bioinformatic methods described herein that target human CCR7 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 17] SEQ ID NOS:518-534 are crRNA sequences generated by the bioinformatic methods described herein that target human CRCP to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 18]SEQ ID NOS:535-551 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCL1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 19] SEQ ID NOS: 552-568 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCL2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 20] SEQ ID NOS: 569-585 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCL3 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 21] SEQ ID NOS:586-602 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCL5 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 22] SEQ ID NOS:603-619 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCL6 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 23]SEQ ID NOS: 620-636 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCL8 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 24] SEQ ID NOS: 637-655 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCR1 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 25] SEQ ID NOS: 656-672 are crRNA sequences generated by the bioinformatic methods described herein that target human CXCR2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 26] SEQ ID NOS:673-720 are crRNA sequences generated by the bioinformatic methods described herein that target human FGF2 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 27] SEQ ID NOS:721-768 are crRNA sequences generated by the bioinformatic methods described herein that target human FGFR1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 28]SEQ ID NOS:769-786 are crRNA sequences generated by the bioinformatic methods described herein that target human IL1A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 29] SEQ ID NOS:787-805 are crRNA sequences generated by the bioinformatic methods described herein that target human IL1B to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 30] SEQ ID NOS:806-839 are crRNA sequences generated by the bioinformatic methods described herein that target human IL1R1 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 31] SEQ ID NOS:840-887 are crRNA sequences generated by the bioinformatic methods described herein that target human IL1RAP to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 32] SEQ ID NOS:888-911 are crRNA sequences generated by the bioinformatic methods described herein that target human IL4 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 33]SEQ ID NOS: 912-928 are crRNA sequences generated by the bioinformatic methods described herein that target human IL6 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 34] SEQ ID NOS: 929-963 are crRNA sequences generated by the bioinformatic methods described herein that target human IL6R to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 35] SEQ ID NOS: 964-990 are crRNA sequences generated by the bioinformatic methods described herein that target human IL6ST to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 36] SEQ ID NOS: 991-1007 are crRNA sequences generated by the bioinformatic methods described herein that target human IL10 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 37] SEQ ID NOS: 1008-1055 are crRNA sequences generated by the bioinformatic methods described herein that target human IL10RA to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 38]SEQ ID NOS: 1056-1082 are crRNA sequences generated by the bioinformatic methods described herein that target human IL10RB to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 39] SEQ ID NOS: 1083-1104 are crRNA sequences generated by the bioinformatic methods described herein that target human IL13 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 40] SEQ ID NOS: 1105-1130 are crRNA sequences generated by the bioinformatic methods described herein that target human IL13RA1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 41] SEQ ID NOS: 1131-1147 are crRNA sequences generated by the bioinformatic methods described herein that target human IL13RA2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 42] SEQ ID NOS: 1148-1173 are crRNA sequences generated by the bioinformatic methods described herein that target human IL17A and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 43]SEQ ID NOS: 1174-1221 are crRNA sequences generated by the bioinformatic methods described herein that target human IL17RA to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 44] SEQ ID NOS: 1222-1238 are crRNA sequences generated by the bioinformatic methods described herein that target human IL18 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing sites and scores, and summarizes several predicted performance indicators. [Figure 45] SEQ ID NOS: 1239-1262 are crRNA sequences generated by the bioinformatic methods described herein that target human IL18R1 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 46] SEQ ID NOS: 1263-1310 are crRNA sequences generated by the bioinformatic methods described herein that target human IL18RAP to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 47] SEQ ID NOS: 1311-1343 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP1 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 48]SEQ ID NOS: 1344-1391 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 49] SEQ ID NOS: 1392-1417 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP3 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 50] SEQ ID NOS: 1418-1436 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP7 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 51] SEQ ID NOS: 1437-1474 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP8 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 52] SEQ ID NOS: 1475-1497 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP10 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 53]SEQ ID NOS: 1498-1541 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP12 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 54] SEQ ID NOS: 1542-1568 are crRNA sequences generated by the bioinformatic methods described herein that target human MMP13 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 55] SEQ ID NOS: 1569-1585 are crRNA sequences generated by the bioinformatic methods described herein that target human MRGPRX2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 56] SEQ ID NOS: 1586-1628 are crRNA sequences generated by the bioinformatic methods described herein that target human NGF to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 57] SEQ ID NOS: 1629-1676 are crRNA sequences generated by the bioinformatic methods described herein that target human NGFR to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 58]SEQ ID NOs: 1677-1724 are crRNA sequences generated by the bioinformatic methods described herein that target human NTF3 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 59] SEQ ID NOS: 1725-1746 are crRNA sequences generated by the bioinformatic methods described herein that target human NTF4 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 60] SEQ ID NOS: 1747-1794 are crRNA sequences generated by the bioinformatic methods described herein that target human NTRK1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 61] SEQ ID NOS: 1795-1842 are crRNA sequences generated by the bioinformatic methods described herein that target human NTRK2 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 62] SEQ ID NOS: 1843-1859 are crRNA sequences generated by the bioinformatic methods described herein that target human RAMP1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 63]SEQ ID NOS: 1860-1907 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN1A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 64] SEQ ID NOS: 1908-1955 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN2A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 65] SEQ ID NOS: 1956-2003 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN3A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and a score, summarizing several predicted performance indicators. [Figure 66] SEQ ID NOS: 2004-2051 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN4A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 67] SEQ ID NOS: 2052-2099 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN5A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 68]SEQ ID NOS: 2100-2147 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN8A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 69] SEQ ID NOs: 2148-2195 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN9A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 70] SEQ ID NOS: 2196-2243 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN10A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 71] SEQ ID NOS: 2244-2291 are crRNA sequences generated by the bioinformatic methods described herein that target human SCN11A and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 72] SEQ ID NOS: 2292-2308 are crRNA sequences generated by the bioinformatic methods described herein that target human TAC1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 73]SEQ ID NOS: 2309-2325 are crRNA sequences generated by the bioinformatic methods described herein that target human TAC3 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 74] SEQ ID NOS: 2326-2373 are crRNA sequences generated by the bioinformatic methods described herein that target human TACR1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 75] SEQ ID NOS:2374-2421 are crRNA sequences generated by the bioinformatic methods described herein that target human TACR2 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 76] SEQ ID NOS:2422-2469 are crRNA sequences generated by the bioinformatic methods described herein that target human TACR3 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 77] SEQ ID NOS:2470-2509 are crRNA sequences generated by the bioinformatic methods described herein that target human TIMP1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 78]SEQ ID NOS:2510-2557 are crRNA sequences generated by the bioinformatic methods described herein that target human TIMP3 and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 79] SEQ ID NOS: 2558-2574 are crRNA sequences generated by the bioinformatic methods described herein that target human TNF-α and modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 80] SEQ ID NOS:2575-2622 are crRNA sequences generated by the bioinformatic methods described herein that target human TNFRSF1A to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 81] SEQ ID NOS: 2623-2670 are crRNA sequences generated by the bioinformatic methods described herein that target human TNFRSF1B to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 82] SEQ ID NOs: 2671-2718 are crRNA sequences generated by the bioinformatic methods described herein that target human YAP1 to modify and / or ablate expression of its encoded product. Additional information includes chromosomal genomic coordinates (assembly hg38) of the editing site and scores, and summarizes several predicted performance indicators. [Figure 83A]These figures collectively illustrate the results of cell-based and in silico gene editing analyses of crRNA sequences targeting the (A) hIL1A, (B) hIL1B, (C) cIL1A, and (D) cIL1B genes. "o" indicates the CRISPR cleavage position within the amino acid (AA) translational frame. "*" indicates the optimization score from Doench, Fusi et al. (2016). This score is optimized for 20-bp guides containing an NGG PAM. Scores range from 0 to 100; higher is better. **Specificity score from Hsu et al. (2013). Scores range from 0 to 100; higher is better. ***This score is based on experiments in U2OS. A high-precision score (>0.4) suggests that DNA repair outcomes are uniform and concentrated in only a handful of unique genotypes. ****This score is based on experiments in U2OS. High (>80%) frameshift frequencies tend to place protein-coding genes out of frame. Because 1-bp insertions and 1-2 bp deletions are particularly common repair outcomes, typical genomic frameshift frequencies are greater than 66%. ^ Composite score = (off-target score + precision score * 100 + frameshift) / 3. † Pipe symbol "|" indicates CRISPR cut site. Curly brackets "{}" indicate insertions. Hyphen "-" indicates deletions. $ Potential off-target site. Scoring according to Hsu et al. (2013). Off-target sites have a score of 100. [Figure 83B]These figures collectively illustrate the results of cell-based and in silico gene editing analyses of crRNA sequences targeting the (A) hIL1A, (B) hIL1B, (C) cIL1A, and (D) cIL1B genes. "o" indicates the CRISPR cleavage position within the amino acid (AA) translational frame. "*" indicates the optimization score from Doench, Fusi et al. (2016). This score is optimized for 20-bp guides containing an NGG PAM. Scores range from 0 to 100; higher is better. **Specificity score from Hsu et al. (2013). Scores range from 0 to 100; higher is better. ***This score is based on experiments in U2OS. A high-precision score (>0.4) suggests that DNA repair outcomes are uniform and concentrated in only a handful of unique genotypes. ****This score is based on experiments in U2OS. High (>80%) frameshift frequencies tend to place protein-coding genes out of frame. Because 1-bp insertions and 1-2 bp deletions are particularly common repair outcomes, typical genomic frameshift frequencies are greater than 66%. ^ Composite score = (off-target score + precision score * 100 + frameshift) / 3. † Pipe symbol "|" indicates CRISPR cut site. Curly brackets "{}" indicate insertions. Hyphen "-" indicates deletions. $ Potential off-target site. Scoring according to Hsu et al. (2013). Off-target sites have a score of 100. [Figure 83C]These figures collectively illustrate the results of cell-based and in silico gene editing analyses of crRNA sequences targeting the (A) hIL1A, (B) hIL1B, (C) cIL1A, and (D) cIL1B genes. "o" indicates the CRISPR cleavage position within the amino acid (AA) translational frame. "*" indicates the optimization score from Doench, Fusi et al. (2016). This score is optimized for 20-bp guides containing an NGG PAM. Scores range from 0 to 100; higher is better. **Specificity score from Hsu et al. (2013). Scores range from 0 to 100; higher is better. ***This score is based on experiments in U2OS. A high-precision score (>0.4) suggests that DNA repair outcomes are uniform and concentrated in only a handful of unique genotypes. ****This score is based on experiments in U2OS. High (>80%) frameshift frequencies tend to place protein-coding genes out of frame. Because 1-bp insertions and 1-2 bp deletions are particularly common repair outcomes, typical genomic frameshift frequencies are greater than 66%. ^ Composite score = (off-target score + precision score * 100 + frameshift) / 3. † Pipe symbol "|" indicates CRISPR cut site. Curly brackets "{}" indicate insertions. Hyphen "-" indicates deletions. $ Potential off-target site. Scoring according to Hsu et al. (2013). Off-target sites have a score of 100. [Figure 83D]These figures collectively illustrate the results of cell-based and in silico gene editing analyses of crRNA sequences targeting the (A) hIL1A, (B) hIL1B, (C) cIL1A, and (D) cIL1B genes. "o" indicates the CRISPR cleavage position within the amino acid (AA) translational frame. "*" indicates the optimization score from Doench, Fusi et al. (2016). This score is optimized for 20-bp guides containing an NGG PAM. Scores range from 0 to 100; higher is better. **Specificity score from Hsu et al. (2013). Scores range from 0 to 100; higher is better. ***This score is based on experiments in U2OS. A high-precision score (>0.4) suggests that DNA repair outcomes are uniform and concentrated in only a handful of unique genotypes. ****This score is based on experiments in U2OS. High (>80%) frameshift frequencies tend to place protein-coding genes out of frame. Because 1-bp insertions and 1-2 bp deletions are particularly common repair outcomes, typical genomic frameshift frequencies are greater than 66%. ^ Composite score = (off-target score + precision score * 100 + frameshift) / 3. † Pipe symbol "|" indicates CRISPR cut site. Curly brackets "{}" indicate insertions. Hyphen "-" indicates deletions. $ Potential off-target site. Scoring according to Hsu et al. (2013). Off-target sites have a score of 100. [Figure 84A] Collectively, the results of functional assays on edited or control canine chondrocytes measuring (A, B) cIL1A and (C, D) cIL1B release at 6 and 24 hours after exposure to PBS or LPS are illustrated. [Figure 84B] Collectively, the results of functional assays on edited or control canine chondrocytes measuring (A, B) cIL1A and (C, D) cIL1B release at 6 and 24 hours after exposure to PBS or LPS are illustrated. [Figure 84C]Collectively, the results of functional assays on edited or control canine chondrocytes measuring (A, B) cIL1A and (C, D) cIL1B release at 6 and 24 hours after exposure to PBS or LPS are illustrated. [Figure 84D] Collectively, the results of functional assays on edited or control canine chondrocytes measuring (A, B) cIL1A and (C, D) cIL1B release at 6 and 24 hours after exposure to PBS or LPS are illustrated. [Figure 85A] Collectively, the results of functional assays in edited and control chondrocytes measuring (A, B) hIL1A and (C, D) cIL1B release after 6 and 24 hours of exposure to PBS or LPS are illustrated. [Figure 85B] Collectively, the results of functional assays in edited and control chondrocytes measuring (A, B) hIL1A and (C, D) cIL1B release after 6 and 24 hours of exposure to PBS or LPS are illustrated. [Figure 85C] Collectively, the results of functional assays in edited and control chondrocytes measuring (A, B) hIL1A and (C, D) cIL1B release after 6 and 24 hours of exposure to PBS or LPS are illustrated. [Figure 85D] Collectively, the results of functional assays in edited and control chondrocytes measuring (A, B) hIL1A and (C, D) cIL1B release after 6 and 24 hours of exposure to PBS or LPS are illustrated. [Figure 86] 1 illustrates the results of tissue-specific splicing and expression analysis of the hIL1A gene. [Figure 87] 1 illustrates the results of tissue-specific splicing and expression analysis of the hIL1B gene. [Figure 88]This figure illustrates the results of in silico analysis of crRNAs targeting either hIL1A or hIL1B. The on-target score (see Doench et al.) is optimized for a 20-bp gRNA with an NGG protospacer adjacent motif (PAM). Scores range from 0 to 1. The precision score is based on experiments in U2OS cells. A precision score (>0.4) suggests that DNA repair outcomes are uniform and concentrated in only a handful of unique genotypes. The frameshift percentage is based on experiments in U2OS cells. A high (>80%) frameshift frequency tends to shift protein-coding genes out of frame. Because 1-bp insertions and 1-2 bp deletions are particularly common repair outcomes, the typical genome frameshift frequency is greater than 66%. The off-target score from CRISPR assesses the number of matches within the genome with a given number of mismatches. Mismatches in the seed sequence have more deleterious effects. [Figure 89] The results of splicing and functional analyses of the cIL1A and cIL1B genes are shown, using the reference canine genome assembly (CanFam3.1). [Figure 90] Illustrates the results of in silico analysis of crRNAs targeting either the cIL1A or cIL1B gene. [Figure 91A] Collectively illustrate the knockdown efficacy of selected sgRNAs in (A) human chondrocytes, (B) canine chondrocytes, and (C) canine synoviocytes. [Figure 91B] Collectively illustrate the knockdown efficacy of selected sgRNAs in (A) human chondrocytes, (B) canine chondrocytes, and (C) canine synoviocytes. [Figure 91C] Collectively illustrate the knockdown efficacy of selected sgRNAs in (A) human chondrocytes, (B) canine chondrocytes, and (C) canine synoviocytes. [Figure 92] Illustrates the results of an in silico analysis of the off-target effects of multiple sgRNAs in canine cells. [Figure 93A](A) Collectively illustrates the efficacy of enhanced specificity Cas9 (espCas9) to eliminate off-target editing of the indicated sgRNAs in canine cells, compared to (B) the effect with canonical spCas9. [Figure 93B] (A) Collectively illustrates the efficacy of enhanced specificity Cas9 (espCas9) to eliminate off-target editing of the indicated sgRNAs in canine cells, compared to (B) the effect with canonical spCas9. [Figure 94A] Collectively illustrate a summary of the editing activity of crRNAs targeting IL1A and IL1B in (A) human and (B) canine chondrocytes. [Figure 94B] Collectively illustrate a summary of the editing activity of crRNAs targeting IL1A and IL1B in (A) human and (B) canine chondrocytes. [Figure 95] Illustrates the results of co-administering multiple sgRNAs in canine cells, either simultaneously or sequentially. [Figure 96A] Collectively, the sequence alignments of (A) the IL1A gene and (B) the IL1B gene for different mammalian species (human, horse, mouse, and dog) are illustrated. [Figure 96B] Collectively, the sequence alignments of (A) the IL1A gene and (B) the IL1B gene for different mammalian species (human, horse, mouse, and dog) are illustrated. DETAILED DESCRIPTION OF THE INVENTION

[0009] I. Introduction Provided herein are compositions and methods for silencing the translation of one or more proteins in an animal in need thereof to treat a pain-related disease, illness, or condition.

[0010] 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 the transmembrane domain (i.e., generating a soluble decoy receptor). In some embodiments, CRISPR editing results in the removal of the cytoplasmic domain (i.e., generating a membrane-bound decoy receptor). In certain embodiments, compositions and methods are provided for gene editing of FGF2, CCN2, ADAMTS5, MMP1, and / or NGF.

[0011] 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.

[0012] In some embodiments, pain is ameliorated by silencing a pain signaling protein (or its cognate receptor) through CRISPR editing of the gene encoding the protein (or receptor). In some embodiments, CRISPR editing results in the removal of the transmembrane domain of the pain receptor (i.e., the generation of a soluble decoy receptor). In some embodiments, CRISPR editing results in the removal of the cytoplasmic domain of the pain receptor (i.e., the generation of a membrane-bound decoy receptor). In certain embodiments, the compositions and methods involve the use of (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, or NTRK2), (ii) one or more metalloproteases or regulators thereof (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, or TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18, TNF , CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, or IL18RAP), (iv) one or more regulators of neural signaling (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, or ATP1A1), (v) one or more other regulators of neuronal signaling (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), or (vi) a combination of any of (i) to (v) is provided for ameliorating pain.

[0013] 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 disclosure belongs. All patents and publications referenced herein are incorporated by reference in their entirety.

[0014] The term "FGF2 gene" refers to a mammalian gene encoding a fibroblast growth factor 2 polypeptide. Non-limiting examples of FGF2 genes include NCBI Gene ID: 2247 [human], NCBI Gene ID: 403857 [dog], NCBI Gene ID: 100033955 [horse], NCBI Gene ID: 100135772 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by FGF2 genes include UniProt: P09038; NP_001348594.1 [human], XP_038421156.1 [dog], NP_001182150.1 [horse], XP_044911834.1 [cat]), and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by the above-listed genes act as ligands for the FGF receptors FGFR1, FGFR2, FGFR3, and FGFR4, in addition to binding strongly to heparin and integrins. Additionally, FGF2 signaling is thought to influence local pain sensation, at least through its pro-angiogenic activity, and is involved in pain perception associated with at least IVD degeneration and joint pathology. 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).

[0015] The term "FGFR1 gene" refers to a mammalian gene encoding a fibroblast growth factor 1 polypeptide. Non-limiting examples of FGFR1 genes include NCBI Gene ID: 2260 [human], NCBI Gene ID: 100856477 [dog], NCBI Gene ID: 100057614 [horse], NCBI Gene ID: 101086055 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by FGFR1 genes include UniProt: P11362; NP_001167534.1 [human], XP_038545782.1 [dog], XP_023486323.1 [horse], XP_011279822.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by the genes listed above are tyrosine-protein kinases that act as cell surface receptors for fibroblast growth factors. In that role, they play important roles in regulating embryonic development, cell proliferation, differentiation, and migration. 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).

[0016] "The term CCN2 gene refers to a mammalian gene encoding a cell communication network factor 2 polypeptide. Non-limiting examples of CCN2 genes include NCBI Gene ID: 1490 [human], NCBI Gene ID: 476202 [dog], NCBI Gene ID: 100073098 [horse ... ID:101094598 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CCN2 gene include UniProt:P29279;NP_001892.2 [human], XP_038321343.1 [dog], XP_023506869.1 [horse], XP_023110145.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Typically, the proteins encoded by the above-listed genes are mitogens secreted by vascular endothelial cells and are involved in chondrocyte proliferation and differentiation and cell adhesion in many cell types. 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, dog, horse, and cat forms, respectively).

[0017] The term "ADAMTS5 gene" refers to a mammalian gene encoding an ADAM metallopeptidase having a thrombospondin type 1 motif 5 polypeptide. Non-limiting examples of ADAMTS5 genes include NCBI Gene ID: 11096 [human], NCBI Gene ID: 487713 [dog], NCBI Gene ID: 100066005 [horse], NCBI Gene ID: 101085063 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the ADAMTS5 gene include UniProt: Q9UNA0; NP_008969.2 [human], XP_038299214.1 [dog], XP_023485737.1 [horse], XP_023094603.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, family members share several distinct protein modules, including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a thrombospondin type 1 (TS) motif, with individual family members differing in the number of C-terminal TS motifs. ADAMTS5 has two unique C-terminal domains. Upon proteolytic processing to generate the mature enzyme, ADAMTS5 functions as an aggrecanase that cleaves aggrecan, the major proteoglycan of cartilage, and may mediate cartilage destruction in osteoarthritis. 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).

[0018] The term "ADAMTS1 gene" refers to a mammalian gene encoding an ADAM metallopeptidase having a thrombospondin type 1 motif 1 polypeptide. Non-limiting examples of ADAMTS1 genes include NCBI Gene ID: 9510 [human], NCBI Gene ID: 100686153 [dog], NCBI Gene ID: 791251 [horse], NCBI Gene ID: 101085309 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the ADAMTS1 gene include UniProt: Q9UHI8; NP_008919.3 [human], XP_038374156.1 [dog], XP_023485736.1 [horse], XP_019695041.3 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, family members share several distinct protein modules, including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a thrombospondin type 1 (TS) motif, with individual family members differing in the number of C-terminal TS motifs. ADAMTS1 contains two disintegrin loops and three C-terminal TS motifs. The protein has anti-angiogenic activity, functions as an aggrecanase that cleaves aggrecan, the major proteoglycan of cartilage, and may be involved in its turnover and associated with various inflammatory processes. 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).

[0019] The term "MMP1 gene" refers to a mammalian gene encoding a matrix metallopeptidase 1 polypeptide. Non-limiting examples of MMP1 genes include NCBI Gene ID: 4312 [human], NCBI Gene ID: 489428 [dog], NCBI Gene ID: 100033896 [horse], NCBI Gene ID: 101084217 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP1 gene include UniProt: P03956; NP_001139410.1 [human], XP_038521018.1 [dog], NP_001075316.1 [horse], XP_003992365.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, MMP1 is proteolytically processed from a preproprotein to generate the mature protease. This secreted protease degrades interstitial collagens, including types I, II, and III. 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).

[0020] The term "MMP2 gene" refers to a mammalian gene encoding a matrix metallopeptidase 2 polypeptide. Non-limiting examples of MMP2 genes include NCBI Gene ID: 4313 [human], NCBI Gene ID: 403733 [dog], NCBI Gene ID: 100033948 [horse], NCBI Gene ID: 101098838 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by MMP2 genes include UniProt: P08253; NP_001121363.1 [human], XP_038515255.1 [dog], XP_023492775.1 [horse], XP_003998091.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP2 is a gelatinase A, a type IV collagenase, that contains three fibronectin type II repeats in its catalytic site, allowing it to bind denatured types IV and V collagen and elastin. 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).

[0021] The term "MMP3 gene" refers to a mammalian gene encoding a matrix metallopeptidase 3 polypeptide. Non-limiting examples of MMP3 genes include NCBI Gene ID: 4314 [human], NCBI Gene ID: 403733 [dog], NCBI Gene ID: 100034195 [horse], NCBI Gene ID: 493666 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP3 gene include UniProt: P08254; NP_002413.1 [human], NP_001002967.1 [dog], NP_001075964.1 [horse], XP_003992356.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP3 is an enzyme that degrades fibronectin, laminin, collagens III, IV, IX, and X, and cartilage proteoglycans, and is thought to be involved in wound repair, the progression of atherosclerosis, and tumor initiation. 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).

[0022] The term "MMP7 gene" refers to a mammalian gene encoding a matrix metallopeptidase 7 polypeptide. Non-limiting examples of MMP7 genes include NCBI Gene ID: 4316 [human], NCBI Gene ID: 489432 [dog], NCBI Gene ID: 100068985 [horse], NCBI Gene ID: 727698 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP7 gene include UniProt: P09237; NP_002414.1 [human], NP_001229655.1 [dog], XP_001498859.1 [horse], XP_003992352.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP7 is proteolytically processed to generate a mature protease, which, in addition to activating procollagenase, degrades proteoglycans, fibronectin, elastin, and casein. 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).

[0023] The term "MMP8 gene" refers to a mammalian gene encoding a matrix metallopeptidase 8 polypeptide. Non-limiting examples of MMP8 genes include NCBI Gene ID: 4317 [human], NCBI Gene ID: 489429 [dog], NCBI Gene ID: 100069005 [horse], NCBI Gene ID: 101080995 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP8 gene include UniProt: P22894; NP_001291370.1 [human], XP_038521019.1 [dog], XP_005611595.1 [horse], XP_003992354.3 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP8 is an enzyme that degrades interstitial collagen. 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).

[0024] The term "MMP10 gene" refers to a mammalian gene encoding a matrix metallopeptidase 10 polypeptide. Non-limiting examples of MMP10 genes include NCBI Gene ID: 4319 [human], NCBI Gene ID: 100146442 [horse], NCBI Gene ID: 101081247 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP10 gene include UniProt: P09238; NP_002416.1 [human], XP_005614947.1 [horse], XP_003992355.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP10 is an enzyme that degrades fibronectin and gelatins types I, III, IV, and V. 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).

[0025] The term "MMP12 gene" refers to a mammalian gene encoding a matrix metallopeptidase 12 polypeptide. Non-limiting examples of MMP12 genes include NCBI Gene ID: 4321 [human], NCBI Gene ID: 611789 [dog], NCBI Gene ID: 100069047 [horse], NCBI Gene ID: 101084472 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP12 gene include UniProt: P39900; NP_002417.2 [human], NP_001274067.1 [dog], XP_001498924.2 [horse], XP_003992366.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP12 is an enzyme with significant elastinolytic activity and may be involved in tissue damage and remodeling. 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).

[0026] The term "MMP13 gene" refers to a mammalian gene encoding a matrix metallopeptidase 13 polypeptide. Non-limiting examples of MMP13 genes include NCBI Gene ID: 4322 [human], NCBI Gene ID: 403763 [dog], NCBI Gene ID: 100009711 [horse], NCBI Gene ID: 493679 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the MMP13 gene include UniProt: P45452; NP_002418.1 [human], XP_038521017.1 [dog], NP_001075273.1 [horse], XP_023094811.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP13 is an enzyme that degrades various types of collagen and is involved in wound healing, tissue remodeling, cartilage degradation, bone development, bone mineralization, and ossification. 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).

[0027] The term "TIMP1 gene" refers to a mammalian gene encoding a TIMP metallopeptidase inhibitor 1 polypeptide. Non-limiting examples of TIMP1 genes include NCBI Gene ID: 7076 [human], NCBI Gene ID: 403816 [dog], NCBI Gene ID: 100034220 [horse], NCBI Gene ID: 101095886 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TIMP1 gene include UniProt: P01033; NP_003245.1 [human], NP_001003182.1 [dog], XP_023488949.1 [horse], XP_023105059.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Typically, the protein encoded by this gene functions by forming a one-to-one complex with target metalloproteinases, such as collagenase, irreversibly inactivating them through binding to their catalytic zinc cofactor. TIMP1 acts on MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, and MMP16, but not MMP14, and has been shown to act as a growth factor regulating cell differentiation, migration, and cell death. 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 human, canine, equine, and feline forms, respectively).

[0028] The term "TIMP3 gene" refers to a mammalian gene encoding a TIMP metallopeptidase inhibitor 3 polypeptide. Non-limiting examples of TIMP3 genes include NCBI Gene ID: 7078 [human], NCBI Gene ID: 481289 [dog], NCBI Gene ID: 100033947 [horse], NCBI Gene ID: 101091215 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TIMP3 gene include UniProt: P35625; NP_000353.1 [human], NP_001271368.1 [dog], NP_001075339.1 [horse], XP_003989265.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Typically, the protein encoded by this gene complexes with metalloproteinases (such as collagenases) and irreversibly inactivates them by binding to their catalytic zinc cofactor. TIMP3 is known to act on MMP1, MMP2, MMP3, MMP7, MMP9, MMP13, MMP14, and MMP15. 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).

[0029] The term "CXCL1 gene" refers to a mammalian gene encoding a C-X-C motif chemokine ligand 1 polypeptide. Non-limiting examples of CXCL1 genes include NCBI Gene ID: 2919 [human], NCBI Gene ID: 100034121 [horse], NCBI Gene ID: 102901432 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL1 gene include UniProt: P09341; NP_001502.1 [human], NP_001296409.1 [horse], XP_023108817.2 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene has chemotactic activity for neutrophils and may play a role in inflammation. 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).

[0030] The term "CXCL2 gene" refers to a mammalian gene encoding a C-X-C motif chemokine ligand 2 polypeptide. Non-limiting examples of CXCL2 genes include NCBI Gene ID: 2920 [human], NCBI Gene ID: 100233237 [horse], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL2 gene include UniProt: P19875, Q9UPB8; NP_002080.1 [human], NP_001137427.1 [horse], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene has antibacterial functions through its regulation of inflammatory and immunoregulatory processes. CXCL2 is expressed at sites of inflammation and has been shown to suppress the proliferation of hematopoietic progenitor cells. 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).

[0031] The term "CXCL3 gene" refers to a mammalian gene encoding a C-X-C motif chemokine ligand 3 polypeptide. Non-limiting examples of CXCL3 genes include NCBI Gene ID: 2921 [human] and NCBI Gene ID: 100056258 [horse], as well as synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the CXCL3 gene include UniProt: P19876, Q4W5H9; NP_002081.2 [human], NP_001137265.1 [horse], as well as sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Typically, the protein encoded by this gene is a secreted growth factor that signals through the G protein-coupled receptor CXCR2 and plays a role in inflammation and as a chemoattractant for neutrophils. 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).

[0032] The term "CXCL5 gene" refers to a mammalian gene encoding a C-X-C motif chemokine ligand 5 polypeptide. Non-limiting examples of the CXCL5 gene include NCBI Gene ID: 6374 [human] and its synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL5 gene include UniProt: P19876, Q4W5H9; NP_002081.2 [human], UniProt: P97885 [rat], UniProt: P50228 [mouse], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Normatively, the protein encoded by this gene is thought to interact with the G protein-coupled receptor CXCR2 to promote angiogenesis, remodel connective tissue, and recruit neutrophils. 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] The term "CXCL6 gene" refers to a mammalian gene encoding a C-X-C motif chemokine ligand 6 polypeptide. Non-limiting examples of CXCL6 genes include NCBI Gene ID: 6372 [human], NCBI Gene ID: 106557449 [dog], NCBI Gene ID: 100033988 [horse], NCBI Gene ID: 101094593 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL6 gene include UniProt: P80162; NP_002984.1 [human], XP_038541813.1 [dog], NP_001075355.2 [horse], XP_003985379.3 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene is a chemoattractant for neutrophils and exhibits antibacterial activity. 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).

[0034] The term "CXCL8 gene" refers to a mammalian gene encoding a C-X-C motif chemokine ligand 8 polypeptide. Non-limiting examples of CXCL8 genes include NCBI Gene ID: 3576 [human], NCBI Gene ID: 403850 [dog], NCBI Gene ID: 100037400 [horse], and NCBI Gene ID: 493836 [cat], as well as synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the CXCL8 gene include UniProt: P10145; NP_000575.1 [human], NP_001003200.1 [dog], NP_001077420.2 [horse], and NP_001009281.1 [cat], as well as sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Typically, the protein encoded by this gene is secreted by monocyte macrophages, neutrophils, eosinophils, T lymphocytes, epithelial cells, and fibroblasts, and functions as a chemotactic factor that guides neutrophils to sites of infection. CXCL8 also participates with other cytokines in the pro-inflammatory signaling cascade. 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).

[0035] The term "CCL2 gene" refers to a mammalian gene encoding a "CC motif chemokine ligand 2" polypeptide. Non-limiting examples of CCL2 genes include NCBI Gene ID: 6347 [human], NCBI Gene ID: 403981 [dog], NCBI Gene ID: 100034136 [horse ... ID:100127112 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CCL2 gene include UniProt:P13500; NP_002973.1 [human], NP_001003297.1 [dog], NP_001075400.1 [horse], XP_003996605.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Typically, the protein encoded by this gene acts as a ligand for CCR2, inducing chemotactic activity in monocytes and basophils (but not neutrophils or eosinophils). In some cases, and simply for the purpose of ambiguity, prefixes are added when referring to specific species of proteins or genes (h, c, e, and f refer to the human, dog, horse, and cat forms, respectively).

[0036] The term "CCL3 gene" refers to a mammalian gene encoding a CC motif chemokine ligand 3 polypeptide. Non-limiting examples of CCL3 genes include NCBI Gene ID: 6348 [human], NCBI Gene ID: 448787 [dog], NCBI Gene ID: 100057909 [horse], NCBI Gene ID: 100302540 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CCL3 gene include UniProt: P10147; NP_002974.1 [human], NP_001005251.2 [dog], NP_001108413.1 [horse], NP_001157129.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene plays a role in the inflammatory response through binding to the receptors CCR1, CCR4, and CCR5. 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).

[0037] The term "CCL5 gene" refers to a mammalian gene encoding a CC motif chemokine ligand 5 polypeptide. Non-limiting examples of CCL5 genes include NCBI Gene ID: 6352 [human], NCBI Gene ID: 403522 [dog], NCBI Gene ID: 100033925 [horse], NCBI Gene ID: 493689 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CCL5 gene include UniProt: P13501; NP_001265665.1 [human], NP_001003010.1 [dog], NP_001075332.1 [horse], NP_001009827.1 [cat]), and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene functions as a chemoattractant for blood monocytes, memory T helper cells, and eosinophils, induces the release of histamine from basophils, and activates eosinophils. 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).

[0038] The term "CCL7 gene" refers to a mammalian gene encoding a CC motif chemokine ligand 7 polypeptide. Non-limiting examples of CCL7 genes include NCBI Gene ID: 6354 [human], NCBI Gene ID: 491148 [dog], NCBI Gene ID: 100071714 [horse], NCBI Gene ID: 101096931 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CCL7 gene include UniProt: P80098; NP_006264.2 [human], NP_001010960.1 [dog], XP_005597638.1 [horse], XP_044900774.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene is a secreted chemokine that attracts macrophages during inflammation and metastasis and is an in vivo substrate for MMP2. 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).

[0039] The term "CCL20 gene" refers to a mammalian gene encoding a CC motif chemokine ligand 20 polypeptide. Non-limiting examples of CCL20 genes include NCBI Gene ID: 6364 [human], NCBI Gene ID: 448790 [dog], NCBI Gene ID: 100629808 [horse], NCBI Gene ID: 101089032 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the CCL20 gene include UniProt: P78556; NP_001123518.1 [human], NP_001005254.1 [dog], XP_003365179.2 [horse], XP_003991274.2 [cat], and sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Canonically, the protein encoded by this gene is involved in inflammatory processes and exhibits lymphocyte chemotactic activity. 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).

[0040] The term "CXCR1 gene" refers to a mammalian gene encoding a C-X-C motif chemokine receptor 1 polypeptide. Non-limiting examples of CXCR1 genes include NCBI Gene ID: 3577 [human], NCBI Gene ID: 478906 [dog], NCBI Gene ID: 100058291 [horse], NCBI Gene ID: 101085650 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the CXCR1 gene include UniProt: P25024; NP_000625.1 [human], XP_038303849.1 [dog], XP_001491062.1 [horse], XP_011283865.1 [cat], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the protein encoded by this gene is a receptor for IL8, transducing signals to mediate neutrophil migration to sites of inflammation, among other activities. 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).

[0041] The term "CXCR2 gene" refers to a mammalian gene encoding a C-X-C motif chemokine receptor 2 polypeptide. Non-limiting examples of CXCR2 genes include NCBI Gene ID: 3579 [human], NCBI Gene ID: 478905 [dog], NCBI Gene ID: 100055552 [horse], NCBI Gene ID: 101085396 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the CXCR2 gene include, for example, UniProt: P25025; NP_001161770.1 [human], NP_001003151.2 [dog], XP_005610662.1 [horse], XP_044890398.1 [cat], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the protein encoded by this gene is a receptor for IL8, transducing signals to mediate neutrophil migration to sites of inflammation, among other activities. 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).

[0042] The term "CCR7 gene" refers to a mammalian gene encoding a CC motif chemokine receptor 7 polypeptide. Non-limiting examples of CCR7 genes include NCBI Gene ID: 1236 [human], NCBI Gene ID: 491011 [dog], NCBI Gene ID: 100067673 [horse], NCBI Gene ID: 101084327 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CCR7 gene include UniProt: P32248; NP_001288643.1 [human], XP_038403305.1 [dog], XP_001500231.1 [horse], XP_003996882.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene controls the migration of memory T cells to inflamed tissues and stimulates the maturation of dendritic cells. Signals mediated by this receptor may also function in the pathogenesis of chronic inflammation. 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).

[0043] The term "ADAM17 gene" refers to a mammalian gene encoding an ADAM metallopeptidase domain 17 polypeptide. Non-limiting examples of ADAM17 genes include NCBI Gene ID: 6868 [human], NCBI Gene ID: 475662 [dog], NCBI Gene ID: 100072496 [horse], NCBI Gene ID: 101089004 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the ADAM17 gene include UniProt: P78536; NP_001369706.1 [human], NP_001273795.1 [dog], NP_001295481.1 [horse], XP_003984558.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are proteolytically processed to generate mature proteases, which function by shedding the ectodomain of tumor necrosis factor alpha, thereby releasing soluble tumor necrosis factor alpha from its membrane-bound precursor. 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).

[0044] The term "TNF gene" refers to a mammalian gene encoding a tumor necrosis factor polypeptide. Non-limiting examples of TNF genes include NCBI Gene ID: 7124 [human], NCBI Gene ID: 403922 [dog], NCBI Gene ID: 100033834 [horse], NCBI Gene ID: 493755 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by TNF genes include UniProt: P01375; NP_000585.2 [human], NP_001003244.4 [dog], NP_001075288.2 [horse], NP_001009835.1 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the proteins encoded by these genes are multifunctional pro-inflammatory cytokines that are secreted primarily by macrophages and can bind to (and therefore function through) their receptors, TNFRSF1A and TNFRSF1B. 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).

[0045] The term "TNFRSF1A gene" refers to a mammalian gene encoding a tumor necrosis factor receptor 1 polypeptide. Non-limiting examples of the TNFRSF1A gene include NCBI Gene ID: 7132 [human], NCBI Gene ID: 403634 [dog], NCBI Gene ID: 100059548 [horse], NCBI Gene ID: 493957 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TNFRSF1A gene include UniProt: P19438; NP_001056.1 [human], XP_038295153.1 [dog], XP_023498787.1 [horse], NP_001009361.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by the genes listed above are transmembrane receptor proteins that can bind to their primary ligands, tumor necrosis factor alpha (TNFA) or lymphotoxin alpha (LTA). Upon binding to TNFA, the receptors trimerize, become activated, and initiate 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 human, canine, equine, and feline forms, respectively).

[0046] The term "TNFRSF1B gene" refers to a mammalian gene encoding a tumor necrosis factor receptor 2 polypeptide. Non-limiting examples of TNFRSF1B genes include NCBI Gene ID: 7133 [human], NCBI Gene ID: 487437 [dog], NCBI Gene ID: 100055840 [horse], NCBI Gene ID: 101080392 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TNFRSF1B gene include UniProt: P20333; XP_011540362.1 [human], XP_038387905.1 [dog], XP_023491528.1 [horse], XP_023113905.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by the genes listed above are transmembrane receptor proteins that can bind 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 human, canine, equine, and feline forms, respectively).

[0047] The term "IL4 gene" refers to a mammalian gene encoding an interleukin-4 polypeptide. Non-limiting examples of IL4 genes include NCBI Gene ID: 3565 [human], NCBI Gene ID: 403785 [dog], NCBI Gene ID: 100034225 [horse], NCBI Gene ID: 751514 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL4 genes include UniProt: P05112; NP_000580.1 [human], NP_001003159.1 [dog], NP_001075988.1 [horse], NP_001036804.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are pleiotropic cytokines produced by activated T cells and are considered important cytokines for tissue repair, counteracting the effects of pro-inflammatory type 1 cytokines, but which also promote allergic airway inflammation and mediate acute inflammation, among other activities. 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).

[0048] The term "IL4R gene" refers to a mammalian gene encoding an interleukin-4 receptor polypeptide. Non-limiting examples of IL4R genes include NCBI Gene ID: 3566 [human], NCBI Gene ID: 489957 [dog], NCBI Gene ID: 791252 [horse], NCBI Gene ID: 101096277 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL4R genes include UniProt: P24394; NP_000409.1 [human], NP_001003159.1 [dog], XP_005598791.2 [horse], XP_023102076.2 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are type I transmembrane proteins that, among other activities, can bind interleukin 4 and interleukin 13, regulate IgE production, and promote Th2 cell differentiation. 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).

[0049] The term "IL6 gene" refers to a mammalian gene encoding an interleukin 6 polypeptide. Non-limiting examples of IL6 genes include NCBI Gene ID: 3569 [human], NCBI Gene ID: 403985 [dog], NCBI Gene ID: 100034196 [horse], NCBI Gene ID: 493687 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the IL6 gene include UniProt: P05231; NP_000591.1 [human], NP_001003301.1 [dog], NP_001075965.2 [horse], NP_001009211.2 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the proteins encoded by these genes are cytokines that function in inflammation and B cell maturation, primarily produced at sites of acute and chronic inflammation, where they are secreted into the serum and induce transcriptional inflammatory responses through the interleukin 6 receptor. 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).

[0050] The term "IL6R gene" refers to a mammalian gene encoding an interleukin-6 receptor polypeptide. Non-limiting examples of IL6R genes include NCBI Gene ID: 3560 [human], NCBI Gene ID: 612271 [dog], NCBI Gene ID: 102148787 [horse], and NCBI Gene ID: 101085689 [cat], as well as synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL6R gene include UniProt: P08887; CAA41231.1 [human], XP_038527979.1 [dog], XP_023496854.1 [horse], and XP_023103841.2 [cat], as well as sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Canonically, the proteins encoded by the genes listed above are transmembrane proteins capable of binding to its natural ligand, interleukin-6. This binding event triggers intracellular signaling events that result in a pro-inflammatory 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).

[0051] The term "IL6ST gene" refers to a mammalian gene encoding an interleukin-6 cytokine family signal transducer polypeptide. Non-limiting examples of IL6ST genes include NCBI Gene ID: 3572 [human], NCBI Gene ID: 403545 [dog], NCBI Gene ID: 100051700 [horse], NCBI Gene ID: 101089832 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the IL6ST gene include UniProt: P40189; NP_001177910.1 [human], NP_001273950.1 [dog], XP_023481030.1 [horse], XP_011281205.1 [cat]), and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. 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).

[0052] The term "IL10 gene" refers to a mammalian gene encoding an interleukin-10 polypeptide. Non-limiting examples of IL10 genes include NCBI Gene ID: 3586 [human], NCBI Gene ID: 403628 [dog], NCBI Gene ID: 100034187 [horse], NCBI Gene ID: 493683 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL10 gene include UniProt: P22301; NP_000563.1 [human], NP_001003077.1 [dog], NP_001075959.1 [horse], NP_001009209.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are pleiotropic cytokines that regulate inflammation through binding to their heterodimeric receptors composed of IL10RA and IL10RB, acting on many immune cell types and thereby activating downstream signaling cascades such as the JAK-STAT pathway. 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).

[0053] The term "IL10RA gene" refers to a mammalian gene encoding an interleukin-10 receptor alpha polypeptide. Non-limiting examples of IL10RA genes include NCBI Gene ID: 3587 [human], NCBI Gene ID: 610823 [dog], NCBI Gene ID: 100071172 [horse], NCBI Gene ID: 101087601 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL10RA gene include UniProt: Q13651; NP_001549.2 [human], XP_038520677.1 [dog], XP_014596783.1 [horse], XP_003992449.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are regulators of pro-inflammatory signaling through binding of their ligand IL-10 upon heterodimerization with IL10RB. 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).

[0054] The term "IL10RB gene" refers to a mammalian gene encoding an interleukin-10 receptor beta polypeptide. Non-limiting examples of IL10RB genes include NCBI Gene ID: 3588 [human], NCBI Gene ID: 478404 [dog], NCBI Gene ID: 100052549 [horse], NCBI Gene ID: 101090038 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL10RB gene include UniProt: Q08334; NP_000619.3 [human], XP_038299308.1 [dog], XP_023485821.1 [horse], XP_003991512.2 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are regulators of pro-inflammatory signaling through binding of their ligand IL-10 upon heterodimer formation with IL10RA. 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).

[0055] The term "IL13 gene" refers to a mammalian gene encoding an interleukin-13 polypeptide. Non-limiting examples of IL13 genes include NCBI Gene ID: 3596 [human], NCBI Gene ID: 442990 [dog], NCBI Gene ID: 100034113 [horse], NCBI Gene ID: 101084678 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL13 gene include UniProt: P35225; NP_001341920.1 [human], NP_001003384.1 [dog], NP_001137263.1 [horse], NP_001009209.1 [cat], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the proteins encoded by these genes regulate the production of pro-inflammatory cytokines and chemokines. 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).

[0056] The term "IL13RA1 gene" refers to a mammalian gene encoding an interleukin-13 receptor alpha 1 polypeptide. Non-limiting examples of IL13RA1 genes include NCBI Gene ID: 3597 [human], NCBI Gene ID: 403623 [dog], NCBI Gene ID: 100055312 [horse], NCBI Gene ID: 101091351 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL13RA1 gene include UniProt: P78552; NP_001551.1 [human], XP_038306633.1 [dog], XP_023490026.1 [horse], XP_023104651.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are low-affinity binding partners of IL13 and, in conjunction with IL13RA2, comprise functional receptors. Upon binding to IL13, the receptor complex stimulates the production of pro-inflammatory cytokines and chemokines. 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).

[0057] The term "IL13RA2 gene" refers to a mammalian gene encoding an interleukin-13 receptor alpha 2 polypeptide. Non-limiting examples of IL13RA2 genes include NCBI Gene ID: 3598 [human], NCBI Gene ID: 403622 [dog], NCBI Gene ID: 100057673 [horse], and NCBI Gene ID: 101100114 [cat], as well as synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL13RA2 gene include UniProt: Q14627; NP_000631.1 [human], NP_001003075.1 [dog], XP_023489189.1 [horse], and XP_044906881.1 [cat], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are high-affinity binding partners of IL13 but lack the cytoplasmic domain. Together with IL13RA1, they form a functional receptor that stimulates the production of pro-inflammatory cytokines and chemokines. 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).

[0058] The term "IL17A gene" refers to a mammalian gene encoding an interleukin-17A polypeptide. Non-limiting examples of IL17A genes include NCBI Gene ID: 3605 [human], NCBI Gene ID: 481837 [dog], NCBI Gene ID: 100034142 [horse], NCBI Gene ID: 101095339 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL17A gene include UniProt: Q16552; NP_002181.1 [human], NP_001159350.1 [dog], NP_001137264.1 [horse], XP_006931878.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are proinflammatory cytokines that activate the NF-kappaB signaling pathway through interaction with its heterodimeric receptor complex of IL17RA and IL17RC, thereby activating the transcription of various chemokines, cytokines, and other factors. 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).

[0059] The term "IL17RA gene" refers to a mammalian gene encoding an interleukin-17 receptor A polypeptide. Non-limiting examples of IL17RA genes include NCBI Gene ID: 23765 [human], NCBI Gene ID: 486759 [dog], NCBI Gene ID: 100055511 [horse], NCBI Gene ID: 101095588 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL17RA gene include UniProt: Q96F46; NP_001276834.1 [human], XP_038295433.1 [dog], XP_005610881.1 [horse], XP_023112364.2 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are transmembrane proteins that bind with low affinity to IL17A as part of a multimeric receptor complex. Through its ligand, IL17RA is involved in many inflammatory conditions. 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).

[0060] The term "IL18 gene" refers to a mammalian gene encoding an interleukin-18 polypeptide. Non-limiting examples of IL18 genes include NCBI Gene ID: 3606 [human], NCBI Gene ID: 403796 [dog], NCBI Gene ID: 100034216 [horse], NCBI Gene ID: 493688 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL18 gene include UniProt: Q14116; NP_001230140.1 [human], XP_038520002.1 [dog], XP_005611483.1 [horse], NP_001009213.2 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are pro-inflammatory cytokines that, when engaged with their receptors and co-receptors, IL18R1 and IL18RAP, regulate inflammatory signaling through the NF-kappa B pathway. 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).

[0061] The term "IL18R1 gene" refers to a mammalian gene encoding an interleukin-18 receptor 1 polypeptide. Non-limiting examples of IL18R1 genes include NCBI Gene ID: 8809 [human], NCBI Gene ID: 611438 [dog], NCBI Gene ID: 100058269 [horse], NCBI Gene ID: 493938 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL18R1 gene include UniProt: Q13478; NP_001269328.1 [human], XP_038536128.1 [dog], XP_023474273.1 [horse], NP_001009863.2 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are essential components for transducing IL18-mediated pro-inflammatory signaling. 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).

[0062] The term "IL18RAP gene" refers to a mammalian gene encoding an interleukin-18 receptor accessory protein polypeptide. Non-limiting examples of IL18RAP genes include NCBI Gene ID: 8807 [human], NCBI Gene ID: 481327 [dog], NCBI Gene ID: 100050212 [horse], NCBI Gene ID: 101084868 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL18RAP gene include UniProt: Q53TU5; NP_001380415.1 [human], XP_038536125.1 [dog], XP_014586460.1 [horse], XP_019682529.2 [cat], and sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Canonically, the proteins encoded by these genes are accessory proteins that enhance IL18-mediated pro-inflammatory signaling. 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).

[0063] The term "NGF gene" refers to a mammalian gene encoding a nerve growth factor polypeptide. Non-limiting examples of NGF genes include NCBI Gene ID: 4803 [human], NCBI Gene ID: 403402 [dog], NCBI Gene ID: 100065669 [horse], NCBI Gene ID: 100144611 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by NGF genes include UniProt: P01138; NP_002497.2 [human], XP_038546347.1 [dog], XP_001496237.2 [horse], XP_044889256.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Typically, this secreted protein is incorporated into a larger complex to form a functional homodimer with nerve growth stimulatory activity. The complex is also involved in regulating the growth and differentiation of sympathetic neurons and certain sensory neurons. 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).

[0064] The term "NGFR gene" refers to a mammalian gene encoding a nerve growth factor receptor polypeptide. Non-limiting examples of NGFR genes include NCBI Gene ID: 4804 [human], NCBI Gene ID: 491071 [dog], NCBI Gene ID: 100069694 [horse], NCBI Gene ID: 101101519 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by NGFR genes include UniProt: P08138; NP_002498.1 [human], XP_038531049.1 [dog], XP_023508464.1 [horse], XP_023099534.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes contain four 40-amino acid repeats in their extracellular domains, with six cysteine ​​residues at conserved positions, followed by a serine / threonine-rich region. This cysteine-rich region contains the nerve growth factor binding domain, which enables signal transduction upon binding. 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).

[0065] The term "NTF3 gene" refers to a mammalian gene encoding a neurotrophin-3 polypeptide. Non-limiting examples of NTF3 genes include NCBI Gene ID: 4908 [human], NCBI Gene ID: 493963 [dog], NCBI Gene ID: 100051839 [horse], NCBI Gene ID: 486731 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the NTF3 gene include UniProt: P20783; NP_001096124.1 [human], XP_038293846.1 [dog], XP_023498780.1 [horse], NP_001009367.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes regulate the survival and differentiation of neurons. 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).

[0066] The term "NTF4 gene" refers to a mammalian gene encoding a neurotrophin-4 polypeptide. Non-limiting examples of NTF4 genes include NCBI Gene ID: 4909 [human], NCBI Gene ID: 611987 [dog], NCBI Gene ID: 100054859 [horse], NCBI Gene ID: 101100428 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the NTF4 gene include UniProt: P34130; NP_001382418.1 [human], NP_001177358.2 [dog], XP_023505846.1 [horse], XP_023101354.2 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are proteolytically processed to a mature form, which can promote neuronal survival through binding of its cognate receptor. Dysregulation of this protein is observed in a variety of neurological disorders. 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).

[0067] The term "NTRK1 gene" refers to a mammalian gene encoding a neurotrophic receptor tyrosine kinase 1 polypeptide. Non-limiting examples of NTRK1 genes include NCBI Gene ID: 4914 [human], NCBI Gene ID: 490404 [dog], NCBI Gene ID: 100064594 [horse], NCBI Gene ID: 101081603 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by NTRK1 genes include UniProt: P04629; NP_001007793.1 [human], XP_038527745.1 [dog], XP_023496742.1 [horse], XP_023103311.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are membrane-bound receptors that, among other functions, bind to neutrophils, signal through the MAPK pathway, and regulate cell differentiation. 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).

[0068] The term "NTRK2 gene" refers to a mammalian gene encoding a neurotrophic receptor tyrosine kinase 2 polypeptide. Non-limiting examples of NTRK2 genes include NCBI Gene ID: 4915 [human], NCBI Gene ID: 484147 [dog], NCBI Gene ID: 100061700 [horse], NCBI Gene ID: 101101347 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by NTRK2 genes include UniProt: Q16620; NP_001007098.1 [human], XP_038510982.1 [dog], XP_023482906.1 [horse], XP_023097987.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are membrane-bound receptors that, among other functions, bind to neutrophils, signal through the MAPK pathway, and regulate cell differentiation. 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).

[0069] The term "BDNF gene" refers to a mammalian gene encoding a brain-derived neurotrophic factor polypeptide. Non-limiting examples of BDNF genes include NCBI Gene ID: 627 [human], NCBI Gene ID: 403461 [dog], NCBI Gene ID: 100009689 [horse], NCBI Gene ID: 493690 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the BDNF gene include UniProt: P23560; NP_001137277.1 [human], NP_001002975.1 [dog], NP_001075256.1 [horse], NP_001009828.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are proteolytically processed to a mature form, which can promote neuronal survival through binding of its cognate receptor. Dysregulation of this protein is observed in a variety of neurological disorders. 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).

[0070] The term "SCN1A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 1 polypeptide. Non-limiting examples of SCN1A genes include NCBI Gene ID: 6323 [human], NCBI Gene ID: 478775 [dog], NCBI Gene ID: 100052059 [horse], NCBI Gene ID: 101081823 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by SCN1A genes include UniProt: P35498; NP_001159435.1 [human], XP_038302870.1 [dog], XP_023478839.1 [horse], XP_019693764.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes mediate voltage-dependent sodium ion permeability of excitable membranes and are involved in the sensory perception of mechanical pain (i.e., activation of somatosensory neurons has been shown to induce pain without neurogenic inflammation). 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).

[0071] The term "SCN2A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 2 polypeptide. Non-limiting examples of SCN2A genes include NCBI Gene ID: 6326 [human], NCBI Gene ID: 478773 [dog], NCBI Gene ID: 100051816 [horse], NCBI Gene ID: 101080472 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by SCN2A genes include UniProt: Q99250; NP_001035232.1 [human], XP_038302857.1 [dog], XP_023478830.1 [horse], XP_023115179.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes mediate voltage-dependent sodium ion permeability of excitable membranes. 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).

[0072] The term "SCN3A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 3 polypeptide. Non-limiting examples of SCN3A genes include NCBI Gene ID: 6328 [human], NCBI Gene ID: 478772 [dog], NCBI Gene ID: 100061941 [horse], NCBI Gene ID: 101082587 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by SCN3A genes include UniProt: Q9NY46; NP_001075145.1 [human], XP_038302852.1 [dog], XP_023478823.1 [horse], XP_019693750.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are subunits of voltage-gated sodium channels and are involved in the propagation of action potentials in neurons and muscle tissue. 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).

[0073] The term "SCN4A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 4 polypeptide. Non-limiting examples of SCN4A genes include NCBI Gene ID: 6328 [human], NCBI Gene ID: 119873250 [dog], NCBI Gene ID: 100049793 [horse], NCBI Gene ID: 101098669 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the SCN4A gene include UniProt: Q9NY46; NP_001075145.1 [human], XP_038531923.1 [dog], NP_001075230.2 [horse], XP_006940553.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by these genes are subunits of voltage-gated sodium channels and are involved in the propagation of action potentials in neurons and muscle tissue. 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).

[0074] The term "SCN5A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 5 polypeptide. Non-limiting examples of SCN5A genes include NCBI Gene ID: 6331 [human], NCBI Gene ID: 403497 [dog], NCBI Gene ID: 100034027 [horse], NCBI Gene ID: 101100994 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by SCN5A genes include UniProt: Q14524; NP_000326.2 [human], NP_001002994.1 [dog], NP_001157367.1 [horse], XP_044893792.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are subunits of voltage-gated sodium channels, found primarily in cardiac muscle and involved in the initial upstroke of the action potential in the electrocardiogram. 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).

[0075] The term "SCN8A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 8 polypeptide. Non-limiting examples of SCN8A genes include NCBI Gene ID: 6335 [human], NCBI Gene ID: 477604 [dog], NCBI Gene ID: 100052777 [horse], NCBI Gene ID: 101096578 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the SCN8A gene include UniProt: Q9UQD0; NP_001171455.1 [human], XP_038294063.1 [dog], XP_023499351.1 [horse], XP_023112849.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are ion pore subunits of voltage-gated sodium channels, essential for rapid membrane depolarization during neuronal action potentials. 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).

[0076] The term "SCN9A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 9 polypeptide. Non-limiting examples of SCN9A genes include NCBI Gene ID: 6335 [human], NCBI Gene ID: 100855710 [dog], NCBI Gene ID: 100052120 [horse], NCBI Gene ID: 101082841 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the SCN9A gene include UniProt: Q15858; NP_001352465.1 [human], XP_038302872.1 [dog], XP_023478844.1 [horse], XP_044889827.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are voltage-gated sodium ion channels that have been implicated in a variety of pain disorders, particularly in the development of inflammatory pain. 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).

[0077] The term "SCN10A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 10 polypeptide. Non-limiting examples of SCN10A genes include NCBI Gene ID: 6336 [human], NCBI Gene ID: 477026 [dog], NCBI Gene ID: 100055493 [horse], NCBI Gene ID: 101085569 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the SCN10A gene include UniProt: Q9Y5Y9; NP_001280235.2 [human], NP_001003203.1 [dog], XP_014587037.1 [horse], XP_044893784.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are transmembrane subunits of voltage-gated sodium channels, which may be involved in the development of pain associated with neuropathies. 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).

[0078] The term "SCN11A gene" refers to a mammalian gene encoding a sodium voltage-gated channel alpha 11 polypeptide. Non-limiting examples of SCN11A genes include NCBI Gene ID: 11280 [human], NCBI Gene ID: 485593 [dog], NCBI Gene ID: 100068480 [horse], NCBI Gene ID: 101085312 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the SCN11A gene include UniProt: Q9UI33; NP_001336182.1 [human], XP_038426400.1 [dog], XP_001916634.3 [horse], XP_044893782.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are transmembrane subunits of voltage-gated sodium channels and are highly expressed in pain-sensing neurons of the dorsal root ganglion and trigeminal ganglion. Mutations in the SCN11A gene have been associated with a variety of pain disorders. 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).

[0079] The term "TAC1 gene" refers to a mammalian gene encoding the tachykinin precursor 1 polypeptide. Non-limiting examples of TAC1 genes include NCBI Gene ID: 6863 [human], NCBI Gene ID: 475239 [dog], NCBI Gene ID: 100052324 [horse], NCBI Gene ID: 101095481 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TAC1 gene include UniProt: P20366; NP_003173.1 [human], XP_038541905.1 [dog], XP_014594521.1 [horse], XP_003982840.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are precursors to substance P, neurokinin A, neuropeptide K, and neuropeptide gamma, four products of the tachykinin peptide hormone family. These hormones are thought to function as neurotransmitters that interact with neural receptors and smooth muscle cells. 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).

[0080] The term "TAC3 gene" refers to a mammalian gene encoding a tachykinin precursor 3 polypeptide. Non-limiting examples of TAC3 genes include NCBI Gene ID: 6866 [human], NCBI Gene ID: 607315 [dog], NCBI Gene ID: 100052722 [horse], and NCBI Gene ID: 101089368 [cat], as well as synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the TAC3 gene include UniProt: Q9UHF0; NP_001171525.1 [human], UniProt: A0A8I3N7Z8; NP_001362511.2 [dog], XP_023499603.1 [horse], and XP_019690663.1 [cat], as well as sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Canonically, the proteins encoded by these genes are proteolytically processed to produce mature peptides, which are primarily expressed in the central and peripheral nervous systems and function as neurotransmitters. This peptide is a ligand for the neurokinin-3 receptor. These hormones are thought to function as neurotransmitters that interact with neural receptors and smooth muscle cells. 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).

[0081] The term "TACR1 gene" refers to a mammalian gene encoding a tachykinin receptor 1 polypeptide. Non-limiting examples of TACR1 genes include NCBI Gene ID: 6869 [human], NCBI Gene ID: 403815 [dog], NCBI Gene ID: 100053491 [horse], NCBI Gene ID: 101090094 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TACR1 gene include UniProt: P25103; NP_001049.1 [human], NP_001012637.1 [dog], XP_001499730.1 [horse], XP_003984209.1 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the proteins encoded by these genes are receptors for the tachykinin substance P, also called neurokinin 1. TACR1 activates the phosphatidylinositol-calcium second messenger system and can also bind with less affinity to substance K and neuromedin-K. 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).

[0082] The term "TACR2 gene" refers to a mammalian gene encoding a tachykinin receptor 2 polypeptide. Non-limiting examples of TACR2 genes include NCBI Gene ID: 6865 [human], NCBI Gene ID: 489020 [dog], NCBI Gene ID: 100034168 [horse], and NCBI Gene ID: 101094541 [cat], as well as synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the TACR2 gene include UniProt: P21452; NP_001048.2 [human], NP_001012635.1 [dog], XP_001502752.2 [horse], and XP_044896003.1 [cat], as well as sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Canonically, the proteins encoded by these genes are receptors for the tachykinin substance K, also called neurokinin A. TACR2 activates the phosphatidylinositol-calcium second messenger system and can also bind with less affinity to neuromedin-K and substance P. 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).

[0083] The term "TACR3 gene" refers to a mammalian gene encoding a tachykinin receptor 3 polypeptide. Non-limiting examples of TACR3 genes include NCBI Gene ID: 6870 [human], NCBI Gene ID: 403814 [dog], NCBI Gene ID: 100073088 [horse], NCBI Gene ID: 101093603 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the TACR3 gene include UniProt: P29371; NP_001050.1 [human], NP_001091010.1 [dog], XP_023492571.1 [horse], XP_003985169.3 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the proteins encoded by these genes are receptors for the tachykinin neurokinin 3, also called neurokinin B or neuromedin-K. TACR3 activates the phosphatidylinositol-calcium second messenger system and can also bind substance K and substance P with less affinity. 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).

[0084] The term "MRGPRX2 gene" refers to a mammalian gene encoding the MAS-related GPR family member X2 polypeptide. Non-limiting examples of MRGPRX2 genes include NCBI Gene ID: 117194 [human], NCBI Gene ID: 485410 [dog], NCBI Gene ID: 100071950 [horse ... ID:101097092 [cat]), or encoded gene products (e.g., UniProt:Q96LB1; NP_001290544.1 [human], XP_038285538.1 [dog], XP_023501936.1 [horse], XP_003993155.1 [cat], as well as their sequence variants, alternatively spliced ​​isoforms, and various glycoforms thereof. Normatively, the proteins encoded by these genes enable G protein-coupled receptor activity and neuropeptide binding activity and are involved in the positive regulation of mast cell degranulation and cytokinesis. In some cases, and simply for disambiguation, prefixes are added when referring to specific species proteins or genes (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0085] The term "ATP1A1 gene" refers to a mammalian gene encoding the ATPase Na+ / K+ transport subunit alpha 1 polypeptide. Non-limiting examples of ATP1A1 genes include NCBI Gene ID: 476 [human], NCBI Gene ID: 403992 [dog], NCBI Gene ID: 100034139 [horse], NCBI Gene ID: 101083695 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the ATP1A1 gene include UniProt: P05023; NP_000692.2 [human], NP_001376153.1 [dog], NP_001108004.2 [horse], XP_011283388.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are integral membrane protein subunits of complexes involved in establishing and maintaining electrochemical gradients of Na and K ions across cell membranes, which are essential for osmoregulation and electrical excitability of nerves and muscles. In some cases, and simply for disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0086] The term "CALCA gene" refers to a mammalian gene encoding a calcitonin-related polypeptide alpha polypeptide. Non-limiting examples of CALCA genes include NCBI Gene ID: 796 [human], NCBI Gene ID: 403946 [dog], NCBI Gene ID: 100033906 [horse], NCBI Gene ID: 101095582 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by CALCA genes include UniProt: P01258; NP_001029124.1 [human], NP_001300719.1 [dog], NP_001075323.1 [horse], XP_019667660.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, this gene encodes multiple gene products, including calcitonin, calcitonin gene-related peptide, and katacalcin, through tissue-specific alternative RNA splicing of the gene transcript and cleavage of an inactive precursor protein. The protein is involved in calcium regulation, regulates phosphorus metabolism, and functions as a vasodilator, among other functions. 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).

[0087] The term "CALCB gene" refers to a mammalian gene encoding a calcitonin-related polypeptide beta polypeptide. Non-limiting examples of CALCB genes include NCBI Gene ID: 797 [human], NCBI Gene ID: 403415 [dog], NCBI Gene ID: 100034126 [horse], NCBI Gene ID: 101094539 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by CALCB genes include UniProt: P10092; NP_000719.1 [human], NP_001002948.1 [dog], NP_001075397.1 [horse], XP_044894937.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes act as vasodilators and neurotransmitters, among other functions. 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).

[0088] The term "CALCRL gene" refers to a mammalian gene encoding a calcitonin receptor-like receptor polypeptide. Non-limiting examples of CALCRL genes include NCBI Gene ID: 10203 [human], NCBI Gene ID: 488438 [dog], NCBI Gene ID: 100054281 [horse], NCBI Gene ID: 101086333 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by CALCRL genes include UniProt: Q16602; NP_001258680.1 [human], XP_038303202.1 [dog], XP_023477941.1 [horse], XP_011283721.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, proteins encoded by these genes include receptors for CGRP (including RAMP1) and ADM (including RAMP2 / 3), which activate adenylyl cyclase. 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).

[0089] The term "RAMP1 gene" refers to a mammalian gene encoding a receptor activity-modifying protein 1 polypeptide. Non-limiting examples of RAMP1 genes include NCBI Gene ID: 10267 [human], NCBI Gene ID: 607163 [dog], NCBI Gene ID: 100066550 [horse], NCBI Gene ID: 101092133 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by RAMP1 genes include UniProt: O60894; NP_005846.1 [human], XP_038291846.1 [dog], XP_023498460.1 [horse], XP_044890618.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the proteins encoded by these genes are required to transport calcitonin receptor-like receptors (CRLRs) to the cell membrane and function together with the CRLR as CGRP receptors. 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).

[0090] The term "ADM gene" refers to a mammalian gene encoding an adrenomedullin polypeptide. Non-limiting examples of ADM genes include NCBI Gene ID: 133 [human], NCBI Gene ID: 403817 [dog], NCBI Gene ID: 100033857 [horse], NCBI Gene ID: 101087095 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the ADM gene include UniProt: P35318; NP_001115.1 [human], NP_001003183.1 [dog], NP_001157351.1 [horse], XP_044894880.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene is a 52 aa peptide that has several functions, including vasodilation, regulating hormone secretion, and promoting angiogenesis. 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).

[0091] The term "CRCP gene" refers to a mammalian gene encoding a CGRP receptor component polypeptide. Non-limiting examples of CRCP genes include NCBI Gene ID: 27297 [human], NCBI Gene ID: 479705 [dog], NCBI Gene ID: 100061681 [horse], NCBI Gene ID: 101084503 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by CRCP genes include UniProt: O75575; NP_001035737.1 [human], XP_038523718.1 [dog], XP_001493592.3 [horse], XP_044903465.1 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Canonically, the protein encoded by this gene is an accessory protein of the CGRP receptor that regulates CGRP responsiveness in various tissues. 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).

[0092] The term "YAP1 gene" refers to a mammalian gene encoding a Yes1-related protein polypeptide. Non-limiting examples of YAP1 genes include NCBI Gene ID: 10413 [human], NCBI Gene ID: 479465 [dog], NCBI Gene ID: 100068834 [horse], NCBI Gene ID: 101101408 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the YAP1 gene include UniProt: P46937; NP_001123617.1 [human], XP_038521022.1 [dog], XP_023500466.1 [horse], XP_044894121.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. Canonically, the protein encoded by this gene is involved in development, growth, repair, and homeostasis. 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).

[0093] The term "IL1RAP gene" refers to a mammalian gene encoding an interleukin-1 receptor accessory protein polypeptide. Non-limiting examples of ILRAP1 genes include NCBI Gene ID: 3556 [human], NCBI Gene ID: 488126 [dog], NCBI Gene ID: 100068726 [horse], NCBI Gene ID: 101094125 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the IL1RAP gene include UniProt: Q9NPH3; NP_002173.1 [human], XP_038318680.1 [dog], XP_001498597.2 [horse], XP_044893081.1 [cat], and their sequence variants, alternatively spliced ​​isoforms, and various glycoforms. 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).

[0094] The term "IL1R1 gene" refers to a mammalian gene encoding an interleukin-1 receptor type 1 polypeptide. Non-limiting examples of IL1R1 genes include NCBI Gene ID: 3554 [human], NCBI Gene ID: 481328 [dog], NCBI Gene ID: 100009699 [horse], NCBI Gene ID: 101080705 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the ILR1 gene include UniProt: P14778; NP_001307909.1 [human], XP_038536135.1 [dog], NP_001075263.2 [horse], XP_023107327.2 [cat], and sequence variants, isoforms encoded by alternative splicing, and various 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).

[0095] The term "IL1A gene" refers to a mammalian gene encoding an interleukin-1 alpha polypeptide. Non-limiting examples of IL1A genes include NCBI Gene ID: 3552 [human], NCBI Gene ID: 403782 [dog], NCBI Gene ID: 100064969 [horse], NCBI Gene ID: 493944 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL1A genes include UniProt: P01583; NP_000566.3 [human], NP_001003157.2 [dog], NP_001075969.2 [horse], NP_001009351.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by the genes listed above are pro-inflammatory cytokines that signal through interactions with IL1R1 and IL1RAP to activate various pathways, including MAPK, JNK, and NF-kappa B. 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).

[0096] The term "IL1B gene" refers to a mammalian gene encoding an interleukin-1 beta polypeptide. Non-limiting examples of IL1B genes include NCBI Gene ID: 3553 [human], NCBI Gene ID: 403974 [dog], NCBI Gene ID: 100034237 [horse], NCBI Gene ID: 768274 [cat], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the IL1B gene include UniProt: P01584; NP_000567.1 [human], NP_001033060.1 [dog], NP_001075995.1 [horse], NP_001070882.1 [cat], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Canonically, the proteins encoded by the genes listed above are key mediators of the inflammatory response and pyrogens that signal through interactions with IL1R1 and IL1RAP. In the central nervous system (CNS), IL1B has been shown to contribute to inflammatory pain hypersensitivity, among other pathologies. 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).

[0097] 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.

[0098] "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.

[0099] 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.

[0100] "Spinal conditions or disorders" include, but are not limited to, lower back pain, neck pain, discogenic 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.

[0101] "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 extremities. The pain may be idiopathic or may be associated with one or more underlying conditions (diagnosed or undiagnosed), including, but not limited to, degenerative disc disease, chronic inflammation, arthritis, osteoporosis, trauma (e.g., post-surgery), infection (e.g., discospondylitis), neuropathy, musculoskeletal abnormalities (e.g., spondylolisthesis or spinal stenosis or spondylolisthesis), herniated nucleus pulposus (HNP), annular ligament rupture, facet arthritis, nerve root compression, and / or other degenerative disorders.

[0102] "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, degenerative disc disease, rheumatoid arthritis, osteoporosis, fibromyalgia, chronic inflammation, infection (e.g., disc spondylitis), herniated disc, spondylosis, spinal stenosis, cervical compressive myelopathy, whiplash, and / or other disorders.

[0103] 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.

[0104] 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.

[0105] 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.

[0106] 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.

[0107] 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.

[0108] 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.

[0109] 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.

[0110] 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).

[0111] 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.

[0112] 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.

[0113] 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.

[0114] 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.

[0115] 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 assist 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.

[0116] The term "CRISPR RNA" or "crRNA" refers to the portion of an sgRNA molecule that has complementarity to a target nucleic acid.

[0117] 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.

[0118] 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.

[0119] 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.

[0120] As used herein, the terms "a," "an," or "the" are generally intended to cover both the singular and the plural.

[0121] 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.

[0122] 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.

[0123] 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."

[0124] As used herein, the term "delivery" means providing an entity to a destination. For example, delivering a therapeutic and / or prophylactic agent to a subject can involve administering a nanoparticle composition comprising the therapeutic and / or prophylactic agent to the subject (e.g., via intravenous, intramuscular, intradermal, subcutaneous, intraarticular, or intradiscal routes). Administering a nanoparticle composition to a mammal or mammalian cells can involve contacting one or more cells with the nanoparticle composition.

[0125] As used herein, "native" means occurring in nature without artificial assistance.

[0126] As used herein, "PEG lipid" or "PEGylated lipid" refers to a lipid that includes a polyethylene glycol moiety. These lipids may also be referred to as PEG-modified lipids.

[0127] As used herein, a "phospholipid" is a lipid that includes a phosphate moiety and one or more carbon chains, such as an unsaturated fatty acid chain. A phospholipid may include one or more multiple (e.g., double or triple) bonds (e.g., one or more unsaturations). Certain phospholipids can promote fusion with membranes. For example, cationic phospholipids can interact with one or more negatively charged phospholipids in a membrane (e.g., a cell membrane or an intracellular membrane). The fusion of a phospholipid with a membrane can allow one or more components of a lipid-containing composition to pass through the membrane, for example, allowing one or more components to be delivered to a cell.

[0128] Any of the compositions disclosed herein can be administered to non-human subjects, such as laboratory or livestock animals. Non-limiting examples of non-human subjects include laboratory or research animals, pets, wild or livestock animals, livestock, etc., such as dogs, goats, guinea pigs, hamsters, mice, pigs, non-human primates (e.g., gorillas, monkeys, orangutans, lemurs, baboons, etc.), rats, sheep, horses, cows, etc. As used herein, a "lipid component" refers to a component of a nanoparticle composition that contains one or more lipids. For example, the lipid component can include one or more cationic / ionic, PEGylated, structural, or other lipids, such as phospholipids.

[0129] 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.

[0130] 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., Cas9 protein) and at least one isolated or non-naturally occurring guide RNA (e.g., sgRNA) to effect amplification of a nucleic acid sequence (e.g., genomic DNA).

[0131] 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; for example, (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or their regulators (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7 , TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more regulators of neuronal signaling (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1), (v) one or more regulators of cell signaling (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1), (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) a nucleotide substitution that results in truncation, nonsense mutation, or other type of loss-of-function of the encoded product of any gene in (i)-(v), i.e., mRNA or protein combination; for example, a deletion of one or more nucleotides that results in truncation, nonsense mutation, or other type of loss-of-function of the encoded product of one or more FGF2, CCN2, ADAMTS5, MMP1, or NGF genes;For example, deletions resulting in loss of function of the encoded mRNA or protein due to single-, double-, or other frameshift deletions, or premature stop codons; or insertions resulting in truncations, nonsense mutations, or other types of loss of function of the encoded gene product, such as the encoded gene product (i.e., mRNA or protein) of one or more of the FGF2, CCN2, ADAMTS5, MMP1, or NGF genes; for example, single-, double-, or other frameshift insertions, or insertions resulting in premature stop codons, resulting in modification of a target gene or locus in a eukaryotic cell. In some embodiments, the CRISPR / Cas system of the present disclosure provides for modification of a gene and / or the encoded product of a gene 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 unable to initiate intracellular signaling). In some embodiments, the CRISPR / Cas system of the present disclosure is packaged in an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 through LNP240;

[0132] 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.

[0133] 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.

[0134] 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.

[0135] Class 1 CRISPR / Cas systems: The exact components, compositions, and methods for using Class 1 CRISPR / Cas systems to effect targeted nucleic acid sequence alteration vary, but should minimally include a nuclease (selected from at least Type I and Type III), 1) at least one guide RNA selected from sgRNA, 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 mode of action within eukaryotic cells. To this end, compositions, components, and methods among Class 1 components may be considered functionally interchangeable, and the following details, provided solely for illustrative purposes, do not represent an exhaustive list of class members. In some embodiments, the CRISPR / Cas systems of the present disclosure are packaged within any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0136] 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.

[0137] 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.

[0138] 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.

[0139] 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.

[0140] 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.

[0141] 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.

[0142] 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.

[0143] 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, ARCas9 (also referred to herein as AR-Cas9), SpCas9-D1135E, SpCas9-HF1, HypaCas9, HiFiCas9, xCas9-3.6, xCas9-3.7, Sniper-Cas9, evoCas9, SpartaCas, LZ3Cas9, miCas9, and SuperFi-Cas9. Additional examples of Cas9 variants disclosed below are incorporated by reference in their entirety for all purposes. Huang, X., et al. (2022). Cells, 11(14), 2186.

[0144] 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]

[0145] 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.

[0146] 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.

[0147] 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).

[0148] In certain embodiments, the targeted gene is (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or regulators thereof (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18A, IL19A, IL20A, IL21B, IL22A, IL23A, IL24A, IL25A, IL26A, IL27A, IL28A, IL29A, IL30A, IL31A, IL32A, IL33A, IL34A, IL35A, IL36A, IL37A, IL38A, IL39A, IL39B, IL39C, IL39D, IL39E, IL39F, IL39F, IL39F, IL39H, IL39H, IL39F, IL39H, IL39F, IL39H, IL39H, IL39F, IL39H, IL39H, IL39F, IL39H, IL39 , IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more regulators of neural signaling (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1), (v) one or more other regulators of neuronal signaling (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) any combination of the genes in (i)-(v). In some embodiments, any region of a targeted gene (e.g., a promoter region, a 5' untranslated region, a 3' untranslated region, an exon, an intron, or an exon / intron boundary) is targeted by an RNA-guided nuclease to modify the gene. In some embodiments, a non-coding region of a targeted gene (e.g., an enhancer region, a promoter region, an intron, a 5' UTR, a 3' UTR, a polyadenylation signal) is targeted to modify the gene.

[0149] 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.

[0150] 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.

[0151] 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.

[0152] 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.

[0153] 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.

[0154] 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.

[0155] 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.

[0156] 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.

[0157] 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.

[0158] 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).

[0159] 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).

[0160] 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. In some embodiments, the CRISPR components of the present disclosure are encapsulated in any one of the LNP systems described herein, including, but not limited to, LNP systems LNP001 through LNP240.

[0161] 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.

[0162] 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.

[0163] 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.

[0164] Non-limiting examples of genes that can be silenced or inhibited by permanent gene editing via the TALE method include: (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or regulators thereof (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more regulators of neural signaling (e.g., SCN1A, SCN2A, SCN3A, (i) one or more other regulators of neuronal signaling (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) any combination of genes listed in (i)-(v). 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.

[0165] 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.

[0166] 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).

[0167] 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.

[0168] 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.

[0169] Non-limiting examples of genes that can be silenced or inhibited by permanent gene editing via zinc finger technology include: (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or regulators thereof (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL1C, IL1D, IL1E, IL1F, IL1G, IL1H, IL1I, IL1I, IL1IH ... 4, IL6, IL10, IL13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more regulators of neural signaling (e.g., SCN1A, SCN2A, SCN3A , SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1); (v) one or more other regulators of neuronal signaling (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2); and (vi) any combination of genes listed in (i)-(v). Non-limiting examples of genes that can be permanently gene edited via zinc finger technology to enhance their resulting products to function as decoys or dominant negatives include: In one aspect, the present disclosure provides compositions for upregulation of protein receptors (wild-type or gene-edited), including those that bind anti-inflammatory cytokines via zinc finger technology.

[0170] 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.

[0171] 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.

[0172] 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. Some embodiments are further useful when related to encapsulation in an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 through LNP240.

[0173] 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.

[0174] 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.

[0175] 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.

[0176] 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.

[0177] 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.

[0178] 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).

[0179] 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.

[0180] 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).

[0181] 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.

[0182] 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 IL1RAP.

[0183] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of IL1R1 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0184] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hIL1R1 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0185] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cIL1R1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0186] 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, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of eIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of eIL1R1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0187] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fIL1R1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0188] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting IL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fIL1RAP. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of IL1RAP is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0189] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hIL1RAP is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0190] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cIL1RAP is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0191] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eIL1RAP is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0192] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fIL1RAP is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0193] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TGFBR1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0194] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of hTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of hTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of hTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of hTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hTGFBR1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0195] 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, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTGFBR1. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cTGFBR1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0196] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTGFBR1 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0197] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fTGFBR1. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fTGFBR1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0198] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TGFBR2 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0199] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hTGFBR2 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0200] In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 1 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 2 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 3 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 4 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 5 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 6 of cTGFBR2. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 7 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of cTGFBR2 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0201] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTGFBR2 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0202] 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, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTGFBR2. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of fTGFBR2 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0203] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting IL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of IL6R is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0204] 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, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of hIL6R. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of hIL6R is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0205] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cIL6R is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0206] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eIL6R is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0207] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fIL6R is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0208] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting IL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fIL6ST. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of IL6ST is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0209] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 19 of hIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hIL6ST is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0210] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of cIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cIL6ST is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0211] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 15 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 16 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 17 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 18 of eIL6ST. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eIL6ST is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0212] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fIL6ST is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0213] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTNFRSF1A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TNFRSF1A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0214] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hTNFRSF1A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0215] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cTNFRSF1A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0216] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTNFRSF1A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0217] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fTNFRSF1A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0218] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTNFRSF1B. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TNFRSF1B is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0219] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of hTNFRSF1B is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0220] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cTNFRSF1B is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0221] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTNFRSF1B is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0222] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fTNFRSF1B is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0223] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TNFRSF3 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0224] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of hTNFRSF3 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0225] 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 the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cTNFRSF3 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0226] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTNFRSF3 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0227] 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 the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF3. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fTNFRSF3 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0228] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTNFRSF4. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TNFRSF4 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0229] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of hTNFRSF4 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0230] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cTNFRSF4 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0231] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTNFRSF4 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0232] 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. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fTNFRSF4 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0233] In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting TNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting hTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting cTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting eTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting fTNFRSF11A. In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of TNFRSF11A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0234] 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.In some embodiments, a CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting exons of hTNFRSF11A is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0235] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of cTNFRSF11A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0236] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of eTNFRSF11A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0237] 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, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprises one or more sgRNAs targeting exon 11 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for the treatment of OA comprising one or more sgRNAs targeting an exon of fTNFRSF11A is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs comprise any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0238] 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. In some embodiments, a CRISPR gene editing system for the treatment of PsA comprising one or more sgRNAs targeting an exon of IL1R1 is delivered via one or more lipid nanoparticles (LNPs), where the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001 through LNP240.

[0239] 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 the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 20 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of psoriatic arthritis comprises one or more sgRNAs targeting exon 21 of hIL1R1. In some embodiments, a CRISPR gene editing system for the treatment of PsA comprising one or more sgRNAs targeting an exon of hIL1R1 is delivered via one or more lipid nanoparticles (LNPs), wherein the LNPs include any one of the LNP systems described herein, such as, but not limited to, LNP systems LNP001-LNP240.

[0240] 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 m...

Claims

1. A pharmaceutical composition for treating a disorder, wherein the composition is (i) RNA guide nuclease or nucleic acid encoding RNA guide nuclease, (ii) FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2, ADAM17, ADA MTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3 CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10 , IL13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL6ST A pharmaceutical composition comprising a plurality of lipid nanoparticles (LNPs) encapsulating at least one guide RNA or nucleic acid encoding at least one guide RNA that targets a gene selected from IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP, SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1, CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2, TGFB, TGFBR1, and TGFBR2.

2. The pharmaceutical composition according to claim 1, wherein the at least one guide RNA comprises a crRNA sequence selected from any one of sequence numbers 1 to 2731.

3. The pharmaceutical composition according to claim 1, wherein the disorder is a musculoskeletal disorder, neoplasia, neurological disorder, cardiac disorder, inflammatory disorder, digestive disorder, respiratory disorder, renal disorder, metabolic disorder, ocular disorder, or autoimmune disorder.

4. The pharmaceutical composition according to claim 1, wherein the plurality of LNPs contain an ionic lipid, and the ionic lipid is SM-102.

5. The pharmaceutical composition according to claim 1, wherein the plurality of LNPs contain an ionic lipid, and the ionic lipid is ALC-0315.

6. The pharmaceutical composition according to claim 1, wherein the plurality of LNPs contain an ionic lipid, and the ionic lipid is LP-01.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the RNA guide nuclease or the nucleic acid encoding the RNA guide nuclease is either the RNA guide nuclease or the DNA encoding the RNA guide nuclease.

8. The pharmaceutical composition according to any one of claims 1 to 6, wherein the RNA guide nuclease or the nucleic acid encoding the RNA guide nuclease is the mRNA encoding the RNA guide nuclease.

9. The pharmaceutical composition according to any one of claims 1 to 6, wherein the RNA guide nuclease is a Cas protein, and optionally the Cas protein is a Cas9 protein.

10. The Cas9 protein is esCas9, hfCas9, peCas9, and ARC The pharmaceutical composition according to claim 9, which is an S. pyogenes Cas9 polypeptide selected from the group consisting of as9.

11. The pharmaceutical composition according to any one of claims 1 to 6, wherein the at least one guide RNA or nucleic acid encoding the at least one guide RNA is the at least one guide RNA.

12. The pharmaceutical composition according to any one of claims 1 to 6, wherein the at least one guide RNA or nucleic acid encoding the at least one guide RNA is DNA encoding the at least one guide RNA.

13. The pharmaceutical composition according to any one of claims 1 to 6, wherein the at least one guide RNA is a single guide RNA (sgRNA).

14. The pharmaceutical composition according to any one of claims 1 to 6, wherein the at least one guide RNA targets a human gene.

15. The pharmaceutical composition according to any one of claims 1 to 6, wherein the at least one guide RNA targets a canine gene, a horse gene, or a feline gene.

16. The pharmaceutical composition according to any one of claims 1 to 6, wherein the composition is formulated for intra-articular injection within a target joint, for intradiscal injection, for peridiscal injection, for intraspinal injection, for epidural injection, or for injection into the facet joints of the spine.

17. The pharmaceutical composition according to any one of claims 1 to 6, wherein the plurality of LNPs include structural lipids, and the structural lipids are cholesterol.

18. The pharmaceutical composition according to any one of claims 1 to 6, wherein the plurality of LNPs comprises a plurality of particles having a diameter greater than 100 nm.

19. The pharmaceutical composition according to any one of claims 1 to 6, wherein the plurality of LNPs comprises a plurality of particles having a diameter of about 60 nm to about 120 nm.

20. The pharmaceutical composition according to any one of claims 1 to 6, wherein the plurality of LNPs comprises an LNP system selected from any one of LNP001 to LNP240.

21. Use of the pharmaceutical composition according to any one of claims 1 to 6 in the manufacture of a pharmaceutical for treating a disease or disorder in a person who requires treatment for such treatment.

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