Recombinant nucleic acid encoding one or more cosmetic proteins for cosmetic applications

Abstract

The present disclosure provides recombinant nucleic acids comprising one or more polynucleotides encoding one or more cosmetic proteins (e.g., one or more human collagens); a virus comprising the recombinant nucleic acid; compositions (e.g., cosmetic formulations) comprising the recombinant nucleic acid and / or the virus; methods of use thereof; and articles or kits thereof.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Application Serial No. 62 / 663,476, filed on April 27, 2018, which is incorporated herein by reference in its entirety.

[0003] Sequence Listing Submission as ASCII Text File

[0004] The following submissions in ASCII text files are incorporated herein by reference in their entirety: Computer Readable Form (CRF) of a Sequence Listing (File Name: 761342000640SEQLIST.txt, Record Date: April 26, 2019, Size: 437KB). Technical Field

[0005] The present disclosure relates in part to recombinant nucleic acids comprising one or more polynucleotides encoding one or more cosmetic proteins (e.g., one or more human collagen proteins); viruses comprising the recombinant nucleic acids; compositions (e.g., cosmetic preparations) comprising the recombinant nucleic acids and / or the viruses; methods of using them; and their products or kits. Background Art

[0006] Like all organs in the human body, the skin is subjected to continuous and often cumulative changes over time. The aging of the skin occurs due to numerous factors, including inherent changes in the skin, the influence of gravity and the facial muscles acting on the skin, the loss of soft tissue or displacement and tissue elasticity. It is of concern that the "aging" phenotype of the skin can be accelerated by environmental factors, most particularly long-term exposure to ultraviolet radiation (e.g., from the sun). Clinically, the aging phenotype of the skin can be described as wrinkling, sinking, and / or compared to its young counterpart, usually with less elasticity and resilience, but the changes in this phenotype are present between natural sequential aging and photoaging.

[0007] The dermal extracellular matrix (ECM) constitutes the bulk of the skin and imparts both strength and resilience. Collagen, the main component of connective tissue that provides support to the skin, decreases with individual age. In aging skin, collagen fibrils show high levels of degradation and breakage, and are replenished at a reduced rate by dermal fibroblasts. These degraded and broken collagen bundles become loose and lose strength (thus destroying the structural organization of the dermal ECM), and inevitably lead to the "aging" appearance of the skin.

[0008] Numerous skin care products have been developed to improve the appearance of human skin. Wrinkles and skin folds are commonly treated with dermal and subdermal injections of cosmetic facial fillers; however, this superficial approach does not address the structural changes underlying skin aging, particularly the damage or loss of collagen. Therefore, there is a clear need for alternative strategies to supplement, strengthen or replace dermal ECM components (e.g., human collagen) in individuals who want to combat or reverse the physiological effects of skin aging.

[0009] All references cited herein, including patent applications, patent publications, non-patent literature, and NCBI / UniProtKB / Swiss-Prot accession numbers, are incorporated by reference in their entirety to the same extent as if each individual reference was specifically and individually indicated to be incorporated by reference. Summary of the invention

[0010] To meet these and other needs, provided herein are recombinant nucleic acids (e.g., recombinant herpes virus genomes) encoding one or more cosmetic proteins for use in viruses (e.g., herpes viruses), compositions, formulations, medicaments, and / or methods for cosmetic / cosmetic applications (e.g., treatment of wrinkles). The inventors have shown that the recombinant attenuated viruses described herein are capable of 1) effectively transducing human epidermal / dermal cells, and 2) successfully expressing the encoded exogenous human collagen (mRNA and protein), wherein the protein can then be localized to appropriate regions in skin-equivalent organotypic cultures (see, e.g., Example 2). In addition, the inventors have shown that the viruses described herein can be successfully administered topically or intradermally without significant host cell cytotoxicity, thereby allowing human collagen expressed from these viruses to be localized to appropriate regions of the dermal ECM after in vivo administration without observable damage to the skin (see, e.g., Examples 3 and 7). In addition, the inventors have shown that a variety of different HSV backbones can be used to construct viruses that express human collagen (see, e.g., Example 2), a variety of strategies can be used to successfully express more than one human collagen from a single recombinant genome (see, e.g., Example 5), and candidate viruses can successfully express human collagen in multiple relevant in vitro and in vivo models of temporal or UV-induced skin aging (see, e.g., Examples 6 and 7). In addition, the inventors have shown that the viruses described herein can be successfully engineered to express other cosmetic proteins (e.g., human laminin) in vitro and in vivo, where these proteins are localized to appropriate regions of the dermal ECM (see, e.g., Example 8). Without wishing to be bound by theory, the data described herein provide strong evidence that the recombinant nucleic acids and / or viruses disclosed herein can constitute a novel means for delivering cosmetic proteins (e.g., human collagens, such as human collagen 1 and human collagen 3), and in particular for supplementing or replacing natural human dermal ECM proteins (e.g., to reduce the appearance of age- or light-induced wrinkles) in cosmetic applications.

[0011] Therefore, some aspects of the present disclosure relate to a recombinant herpesvirus genome comprising a first polynucleotide encoding a first polypeptide, wherein the first polypeptide comprises a first beauty protein. In some embodiments, the recombinant herpesvirus genome comprises two or more copies of the first polynucleotide. In some embodiments, the recombinant herpesvirus genome is replication-competent. In some embodiments that can be combined with any previous embodiment, the recombinant herpesvirus genome is selected from recombinant herpes simplexvirus genome, recombinant varicella zoster virus genome, recombinant human cytomegalovirus genome, recombinant herpesvirus 6A genome, recombinant herpesvirus 6B genome, recombinant herpesvirus 7 genome, recombinant Kaposi's sarcoma (Kaposi's sarcoma) related herpesvirus genome and any derivatives thereof. In some embodiments that can be combined with any previous embodiment, the recombinant herpesvirus genome is a recombinant herpes simplexvirus genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivative thereof.

[0012] In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation. In some embodiments that can be combined with any previous embodiment, the inactivation mutation is in a herpes simplex virus gene. In some embodiments, the inactivation mutation is the deletion of the coding sequence of a herpes simplex virus gene. In some embodiments, the herpes simplex virus gene is selected from infected cell protein (Infected Cell Protein, ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (thymidine kinase, tk), long unique region (UL) 41 and UL55. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in one or two copies of the ICP4 gene. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in the ICP22 gene. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in the UL41 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in one or two copies of the ICPO gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the junction region. In some embodiments, the recombinant herpes simplex virus genome comprises a deletion in the junction region. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or both of the ICP4 viral loci.

[0013] In some embodiments that may be combined with any of the preceding embodiments, the first cosmetic protein is selected from the group consisting of a first collagen, a first fibronectin, a first elastin, a first lumican protein, a first vitronectin, a first vitronectin receptor protein, a first laminin, a first neuromodulator protein, and a first fibrillin. In some embodiments that may be combined with any of the preceding embodiments, the first cosmetic protein comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21 and 53-64. In some embodiments, the first cosmetic protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the first cosmetic protein is collagen, elastin, fibronectin, or laminin (eg, human collagen, human elastin, human fibronectin, or human laminin). In some embodiments that may be combined with any of the preceding embodiments, the first collagen is human collagen.In some embodiments that may be combined with any of the preceding embodiments, the first collagen is selected from the group consisting of collagen alpha-1 (I) chain polypeptide (COL1-1), collagen alpha-2 (I) chain polypeptide (COL1-2), collagen alpha-1 (II) chain polypeptide (COL2), collagen alpha-1 (III) chain polypeptide (COL3), collagen alpha-1 (IV) chain polypeptide (COL4-1), collagen alpha-2 (IV) chain polypeptide (COL4-2), collagen alpha-3 (IV) chain polypeptide (COL4-3), collagen alpha-4 (IV) chain polypeptide (COL4-4), collagen alpha-5 (IV) chain polypeptide (COL4-5), collagen alpha-6 (IV) chain polypeptide (COL4-6) 、Collagen α-1(V) chain polypeptide (COL5-1), collagen α-2(V) chain polypeptide (COL5-2), collagen α-3(V) chain polypeptide (COL5-3), collagen α-1(VI) chain polypeptide (COL6-1), collagen α-2(VI) chain polypeptide (COL6-2), collagen α-3(VI) chain polypeptide (COL6-3), collagen α-4(VI) chain polypeptide (COL6-4), collagen α-5(VI) chain polypeptide (COL6-5), collagen α-6(VI) chain polypeptide (COL6-6), collagen α-1(VIII) chain polypeptide (COL8), collagen α-1(IX) chain polypeptide (COL9-1), collagen α-2 (IX) chain polypeptide (COL9-2), collagen α-3 (IX) chain polypeptide (COL9-3), collagen α-1 (X) chain polypeptide (COL10), collagen α-1 (XI) chain polypeptide (COL11-1), collagen α-2 (XI) chain polypeptide (COL11-2), collagen α-1 (XII) chain polypeptide (COL12), collagen α-1 (XIII) chain polypeptide (COL13), collagen α-1 (XIV) chain polypeptide (COL14), collagen α-1 (XV) chain polypeptide (COL15), collagen α-1 (XVI) chain polypeptide (COL16), collagen α-1 (XVII) chain polypeptide (COL17), collagen α-1 (X Collagen alpha-1 (XX) chain polypeptide (COL28), collagen alpha-1 (XXV) chain polypeptide (COL29), collagen alpha-1 (XXVIII) chain polypeptide (COL30), collagen alpha-1 (XXVIII) chain polypeptide (COL31), collagen alpha-1 (XXVIII) chain polypeptide (COL32), collagen alpha-1 (XXVIII) chain polypeptide (COL33), collagen alpha-1 (XXVIII) chain polypeptide (COL34), collagen alpha-1 (XXVIII) chain polypeptide (COL35), collagen alpha-1 (XXVI) chain polypeptide (COL36), collagen alpha-1 (XXVII) chain polypeptide (COL37), and collagen alpha-1 (XXVIII) chain polypeptide (COL38).In some embodiments that may be combined with any of the preceding embodiments, the first collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, and COL17. In some embodiments that may be combined with any of the preceding embodiments, the first collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments that may be combined with any of the preceding embodiments, the first collagen is COL3. In some embodiments that may be combined with any of the preceding embodiments, the first collagen is human COL3. In some embodiments that may be combined with any of the preceding embodiments, the first collagen protein comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 17. In some embodiments that may be combined with any of the preceding embodiments, the first cosmetic protein is not collagen alpha-1 (VII) chain polypeptide (COL7).

[0014] In some embodiments, the first polypeptide consists essentially of the first beauty protein. In some embodiments, the first polypeptide consists of the first beauty protein. In some embodiments, the first polypeptide comprises: (a) a first beauty protein; (b) another beauty protein; and (c) a linker polypeptide that connects (a) to (b). In some embodiments, another beauty protein is selected from collagen, fibronectin, elastin, aphrodisiac protein, vitronectin, vitronectin receptor protein, laminin, neuromodulator protein and fibrillin. In some embodiments, another beauty protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, another beauty protein is collagen, elastin, fibronectin or laminin (e.g., human collagen, human elastin, human fibronectin or human laminin). In some embodiments, the other collagen (e.g., another human collagen) is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6 , COL7, COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the other collagen (e.g., another human collagen) is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7, and COL17. In some embodiments, the first cosmetic protein and the other cosmetic protein are different. In some embodiments, the first cosmetic protein is COL1-1 (e.g., human COL1-1), and the other cosmetic protein is COL1-2 (e.g., human COL1-2). In some embodiments, the linker polypeptide is a cleavable linker polypeptide. In some embodiments, the linker polypeptide comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 28-31.

[0015] In some embodiments that may be combined with any of the previous embodiments, the first polynucleotide encodes a polycistronic mRNA comprising: (a) a first open reading frame (ORF) encoding a first polypeptide; (b) a second ORF encoding an additional beauty protein; and (c) an internal ribosome entry site (IRES) separating (a) and (b). In some embodiments, the additional beauty protein is selected from collagen, fibronectin, elastin, aphrodisiac protein, vitronectin, vitronectin receptor protein, laminin, neuromodulator protein, and fibrillin. In some embodiments, the additional beauty protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the additional beauty protein is collagen, elastin, fibronectin, or laminin (e.g., human collagen, human elastin, human fibronectin, or human laminin). In some embodiments, the additional collagen (e.g., additional human collagen) is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6 , COL7, COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the additional collagen (e.g., additional human collagen) is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7, and COL17. In some embodiments, the first cosmetic protein and the additional cosmetic protein are different. In some embodiments, the first cosmetic protein is COL1-1 (e.g., human COL1-1), and the additional cosmetic protein is COL1-2 (e.g., human COL1-2). In some embodiments, the nucleic acid sequence encoding the IRES has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NO: 22 or SEQ ID NO: 23.

[0016] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes virus genome further comprises a second polynucleotide encoding a second beauty protein. In some embodiments, the second beauty protein is selected from collagen, fibronectin, elastin, aphrodisiac protein, vitronectin, vitronectin receptor protein, laminin, neuromodulator protein and fibrillin. In some embodiments, the second beauty protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the second beauty protein is collagen, elastin, fibronectin or laminin (e.g., human collagen, human elastin, human fibronectin or human laminin). In some embodiments, the second collagen (e.g., a second human collagen) is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6 , COL7, COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the second collagen (e.g., a second human collagen) is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7, and COL17. In some embodiments, the first cosmetic protein and the second cosmetic protein are different. In some embodiments, the first cosmetic protein is COL1-1 (e.g., human COL1-1), and the second cosmetic protein is COL1-2 (e.g., human COL1-2). In some embodiments, the first cosmetic protein is COL1-1 (e.g., human COL1-1), and the second cosmetic protein is COL3 (e.g., human COL3).

[0017] In some embodiments that can be combined with any previous embodiment, compared to the corresponding wild-type herpesvirus genome, the recombinant herpesvirus genome has a reduced cytotoxicity when introduced into the target cell. In some embodiments, the target cell is a cell of the epidermis and / or dermis. In some embodiments, the target cell is a human cell. In some embodiments, the target cell is a fibroblast.

[0018] Other aspects of the present disclosure relate to a herpesvirus comprising any recombinant herpesvirus genome as described herein. In some embodiments, herpesvirus is replication-competent. In some embodiments, herpesvirus is replication-defective. In some embodiments, herpesvirus is attenuated. In some embodiments that can be combined with any previous embodiment, compared to corresponding wild-type herpesvirus, herpesvirus has the cytotoxicity of reduction. In some embodiments that can be combined with any previous embodiment, herpesvirus is selected from herpes simplex virus, varicella zoster virus, human cytomegalovirus, herpesvirus 6A, herpesvirus 6B, herpesvirus 7 and Kaposi's sarcoma-associated herpesvirus. In some embodiments that can be combined with any previous embodiment, herpesvirus is herpes simplex virus. In some embodiments, herpes simplex virus is herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2) or any derivative thereof. In some embodiments, herpes simplex virus is herpes simplex virus type 1 (HSV-1).

[0019] Other aspects of the present disclosure relate to a composition comprising: (a) any recombinant herpes virus genome described herein and / or any herpes virus described herein; and (b) an excipient. In some embodiments, the composition is sterile. In some embodiments that can be combined with any previous embodiment, the composition is suitable for topical, transdermal, subcutaneous, intradermal, oral, intranasal, intratracheal, sublingual, buccal, rectal, vaginal, inhalation, intravenous, intraarterial, intramuscular, intracardiac, intraosseous, intraperitoneal, mucosal, intravitreal, subretinal, intraarticular, periarticular, topical or epidermal administration. In some embodiments that can be combined with any previous embodiment, the composition is suitable for intradermal administration. In some embodiments that can be combined with any previous embodiment, the composition is suitable for superficial injection. In some embodiments that can be combined with any previous embodiment, the composition is a cosmetic composition. In some embodiments that can be combined with any previous embodiment, the composition is a skin care product.

[0020] Other aspects of the present disclosure relate to the use of any of the recombinant herpes virus genomes described herein and / or any of the herpes viruses described herein as a medicament (eg, for cosmetic indications).

[0021] Other aspects of the disclosure relate to the use of any of the recombinant herpesvirus genomes described herein and / or any of the herpesviruses described herein as a therapy (eg, as a cosmetic or beauty therapy).

[0022] Other aspects of the present disclosure relate to the use of any of the recombinant herpesvirus genomes described herein and / or any of the herpesviruses described herein for the manufacture of a medicament useful for treating one or more signs or symptoms of dermatological aging.

[0023] Other aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the level of one or more dermal extracellular matrix proteins in a subject, the method comprising administering to the subject an effective amount of any herpes virus described herein and / or any composition described herein.

[0024] Other aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the level of one or more collagens in a subject, the method comprising administering to the subject an effective amount of any herpes virus described herein and / or any composition described herein. In some embodiments, the one or more collagens are collagen 3. In some embodiments, the level of endogenous collagen 3 is reduced due to chronological aging or photoaging.

[0025] Other aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the soft tissue of a subject, the method comprising administering to the subject an effective amount of any herpes virus described herein and / or any composition described herein. In some embodiments, the composition is injected into the soft tissue of the subject.

[0026] Other aspects of the present disclosure relate to a method of improving the skin condition, quality and / or appearance of a subject in need thereof, the method comprising applying to the subject an effective amount of any herpes virus described herein and / or any composition described herein. In some embodiments, the composition is applied to one or more sun damage or other UV exposure sites, rough texture sites, skin depression sites, wrinkle sites, or any combination thereof.

[0027] Other aspects of the present disclosure relate to a method for reducing the appearance of one or more superficial depressions in the skin of a subject in need, the method comprising applying an effective amount of any herpes virus described herein and / or any composition described herein to the subject. In some embodiments, one or more superficial depressions in the skin are selected from the group consisting of: nasolabial folds, crows'feet, frown lines, worry lines, scars, glabellar lines, eyebrow droop, tear troughs, nasal cheek lines, bunny lines, cheek / mid-face droop, marionette lines, poppy dimpling, smile lines, smile lines, chin wrinkles, neck lines, platysma bands, and any combination thereof.

[0028] Other aspects of the present disclosure relate to a method of increasing and / or improving at least one of the texture, smoothness, elasticity, or tone of the skin of a subject in need thereof, the method comprising administering to the subject an effective amount of any herpes virus described herein and / or any composition described herein.

[0029] In some embodiments that may be combined with any of the preceding embodiments, the subject's skin is aging skin. In some embodiments that may be combined with any of the preceding embodiments, the subject's skin has been damaged by exposure to ultraviolet light. In some embodiments that may be combined with any of the preceding embodiments, the subject's skin is wrinkled.

[0030] Other aspects of the present disclosure relate to a method of reducing one or more dermatological signs of aging in a subject in need thereof, the method comprising administering to the subject an effective amount of any herpes virus described herein and / or any composition described herein. In some embodiments, the reduction in one or more dermatological signs of aging is indicated by: (a) treatment, reduction and / or prevention of fine lines and / or wrinkles; (b) reduction in skin pore size; (c) improvement in skin thickness, fullness and / or firmness; (d) improvement in skin smoothness, flexibility and / or softness; (e) improvement in skin tone, radiance and / or clarity; (f) improvement in procollagen and / or collagen production; (g) improvement in skin texture and / or promotion of retexturing; (h) improvement in skin (i) skin radiance and / or brightness is restored; (j) the appearance of skin diminished due to aging and / or menopause is improved; (k) skin moisturization is improved; (l) skin elasticity and / or resilience is increased; (m) skin sagging is treated, reduced and / or prevented; (n) skin firmness is improved; (o) pigment spots, mottled skin and / or scars (such as acne scars) are reduced; (p) the optical properties of the skin in terms of light diffraction or reflection are improved; or (q) any combination thereof.

[0031] In some embodiments that can be combined with any previous embodiment, the experimenter is a person.In some embodiments that can be combined with any previous embodiment, herpesvirus or composition part, transdermal mode, subcutaneous mode, mode on skin, mode in dermis, oral mode, sublingual mode, through buccal mode, through rectal mode, vaginal mode, intravenous mode, intraarterial mode, intramuscular mode, intraosseous mode, intracardiac mode, intraperitoneal mode, through mucosal mode, intravitreal mode, subretinal mode, intraarticular mode, periarticular mode, local mode or by suction to experimenter use.In some embodiments that can be combined with any previous embodiment, herpesvirus or composition are used to experimenter with mode in dermis.In some embodiments that can be combined with any previous embodiment, herpesvirus or composition are used by superficial injection.

[0032] Other aspects of the present disclosure relate to a composition comprising: a herpes simplex virus (HSV) comprising a recombinant nucleic acid, wherein the recombinant nucleic acid comprises a first polynucleotide encoding a first polypeptide, the first polypeptide comprising a first human collagen; and an excipient. In some embodiments, the recombinant nucleic acid comprises two or more copies of the first polynucleotide. In some embodiments that may be combined with any of the previous embodiments, the HSV is replication-deficient. In some embodiments that may be combined with any of the previous embodiments, the HSV is replication-competent. In some embodiments that may be combined with any of the previous embodiments, the HSV is herpes simplex type 1 virus, herpes simplex type 2 virus, or any derivatives thereof.

[0033] In some embodiments, the recombinant nucleic acid is a herpes simplex virus amplicon. In some embodiments, the herpes simplex virus amplicon is an HSV-1 amplicon or an HSV-1 hybrid amplicon. In some embodiments, the HSV-1 hybrid amplicon is an HSV / AAV hybrid amplicon, an HSV / EBV hybrid amplicon, and an HSV / EBV / RV hybrid amplicon, or an HSV / Sleeping Beauty hybrid amplicon.

[0034] In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant HSV-1 genome, a recombinant HSV-2 genome, or any derivative thereof. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in a herpes simplex virus gene. In some embodiments, the herpes simplex virus gene is selected from the group consisting of infected cell protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41 and UL55. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in one or two copies of the ICP4 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP22 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the UL41 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICPO gene. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene. In some embodiments that may be combined with any of the preceding embodiments, the inactivating mutation is a deletion of the coding sequence of one or more genes.

[0035] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in the viral locus. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or two copies of the ICP4 viral locus. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in the ICP22 viral locus. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in the UL41 viral locus. In some embodiments that can be combined with any of the previous embodiments, HSV has reduced cytotoxicity compared to wild-type herpes simplex virus.

[0036] In some embodiments that may be combined with any of the preceding embodiments, the first human collagen is selected from the group consisting of collagen alpha-1 (I) chain polypeptide (COL1-1), collagen alpha-2 (I) chain polypeptide (COL1-2), collagen alpha-1 (II) chain polypeptide (COL2), collagen alpha-1 (III) chain polypeptide (COL3), collagen alpha-1 (IV) chain polypeptide (COL4-1), collagen alpha-2 (IV) chain polypeptide (COL4-2), collagen alpha-3 (IV) chain polypeptide (COL4-3), collagen alpha-4 (IV) chain polypeptide (COL4-4), collagen alpha-5 (IV) chain polypeptide (COL4-5), collagen alpha-6 (IV) chain polypeptide (COL4-6), collagen alpha-1 (III) chain polypeptide (COL4-7), collagen alpha-2 (IV) chain polypeptide (COL4-8), collagen alpha-3 (IV) chain polypeptide (COL4-9), collagen alpha-4 (IV) chain polypeptide (COL4-10), collagen alpha-5 (IV) chain polypeptide (COL4-11), collagen alpha-2 (IV) chain polypeptide (COL4-2), collagen alpha-3 (IV) chain polypeptide (COL4-3), collagen alpha-4 (IV) chain polypeptide (COL4-4), collagen alpha-5 (IV) chain polypeptide (COL4-5), collagen alpha-6 (IV) chain polypeptide (COL4-6), collagen alpha-1 (III) chain polypeptide (COL4-12), collagen alpha-2 (IV) chain polypeptide (COL4-2), collagen alpha-3 (IV) chain polypeptide (COL4-3), collagen alpha-4 (IV) chain polypeptide (COL4-4), collagen alpha-5 (IV) chain polypeptide (COL4-5), collagen alpha-6 (IV) chain polypeptide (COL4-6), collagen alpha -1(V) chain polypeptide (COL5-1), collagen α-2(V) chain polypeptide (COL5-2), collagen α-3(V) chain polypeptide (COL5-3), collagen α-1(VI) chain polypeptide (COL6-1), collagen α-2(VI) chain polypeptide (COL6-2), collagen α-3(VI) chain polypeptide (COL6-3), collagen α-4(VI) chain polypeptide (COL6-4), collagen α-5(VI) chain polypeptide (COL6-5), collagen α-6(VI) chain polypeptide (COL6-6), collagen α-1(VII) chain polypeptide (COL7), collagen α-1(VIII) chain polypeptide (CCOL8), collagen α-1(IX) chain polypeptide (COL6-9), collagen α-2(VI) chain polypeptide (COL6-10), collagen α-3(VI) chain polypeptide (COL6-2), collagen α-3(VI) chain polypeptide (COL6-3), collagen α-4(VI) chain polypeptide (COL6-4), collagen α-5(VI) chain polypeptide (COL6-5), collagen α-6(VI) chain polypeptide (COL6-6), collagen α-1(VII) chain polypeptide (COL7), collagen α-1(VIII) chain polypeptide (CCOL8), collagen α-1(IX) chain polypeptide (COL6-9), collagen α-2 ... L9-1), collagen α-2 (IX) chain polypeptide (COL9-2), collagen α-3 (IX) chain polypeptide (COL9-3), collagen α-1 (X) chain polypeptide (COL10), collagen α-1 (XI) chain polypeptide (COL11-1), collagen α-2 (XI) chain polypeptide (COL11-2), collagen α-1 (XII) chain polypeptide (COL12), collagen α-1 (XIII) chain polypeptide (COL13), collagen α-1 (XIV) chain polypeptide (COL14), collagen α-1 (XV) chain polypeptide (COL15), collagen α-1 (XVI) chain polypeptide (COL16), collagen α-1 (XVII) chain polypeptide (COL17), collagen α-1(XVIII) chain polypeptide (COL18), collagen α-1(XIX) chain polypeptide (COL19), collagen α-1(XX) chain polypeptide (COL20), collagen α-1(XXI) chain polypeptide (COL21), collagen α-1(XXII) chain polypeptide (COL22), collagen α-1(XXIII) chain polypeptide (COL23), collagen α-1(XXIV) chain polypeptide (COL24), collagen α-1(XXV) chain polypeptide (COL25), collagen α-1(XXVI) chain polypeptide (COL26), collagen α-1(XXVII) chain polypeptide (COL27) and collagen α-1(XXVIII) chain polypeptide (COL28).In some embodiments that may be combined with any of the preceding embodiments, the first human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments that may be combined with any of the preceding embodiments, the nucleic acid sequence encoding the first human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments that may be combined with any of the preceding embodiments, the first human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments that may be combined with any of the preceding embodiments, the first human collagen is not COL7.

[0037] In some embodiments that may be combined with any of the preceding embodiments, the first polypeptide comprises: (a) a first human collagen; (b) another human collagen; and (c) a linker polypeptide that connects (a) to (b). In some embodiments, the linker polypeptide is a cleavable linker polypeptide. In some embodiments, the linker polypeptide comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 28-31. In some embodiments, the other human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the other human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments, the nucleic acid sequence encoding the other human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the other human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, the first human collagen and the other human collagen are different.

[0038] In some embodiments that may be combined with any of the preceding embodiments, the first polynucleotide encodes a polycistronic mRNA comprising: (a) a first open reading frame (ORF) encoding a first polypeptide; (b) a second ORF encoding an additional human collagen; and (c) an internal ribosome entry site (IRES) separating (a) and (b). In some embodiments, the nucleic acid sequence encoding the IRES has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleic acid sequence selected from SEQ ID NO: 22 or SEQ ID NO: 23. In some embodiments, the additional human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments, the nucleic acid sequence encoding the additional human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the additional human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, the first human collagen and the additional human collagen are different.

[0039] In some embodiments that may be combined with any of the previous embodiments, the recombinant nucleic acid further comprises a second polynucleotide encoding a second human collagen. In some embodiments, the recombinant nucleic acid comprises two or more copies of the second polynucleotide. In some embodiments, the second human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the second human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments, the nucleic acid sequence encoding the second human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the second human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, the first human collagen and the second human collagen are different.

[0040] In some embodiments that can be combined with any previous embodiment, the recombinant nucleic acid is a recombinant herpes simplex virus genome, and wherein the recombinant herpes simplex virus genome comprises a second polynucleotide in the viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in one or two copies of the ICP4 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in the ICP22 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in the UL41 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or two copies of the ICP4 viral locus, and comprises a second polynucleotide in the ICP22 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or two copies of the ICP4 viral locus, and comprises a second polynucleotide in the UL41 viral locus.

[0041] In some embodiments that may be combined with any of the previous embodiments, the excipient is suitable for transdermal (systemic or topical), transdermal, subcutaneous and / or intradermal administration. In some embodiments that may be combined with any of the previous embodiments, the excipient comprises hydroxypropyl methylcellulose gel. In some embodiments that may be combined with any of the previous embodiments, the excipient is suitable for intradermal administration. In some embodiments that may be combined with any of the previous embodiments, the excipient comprises a phosphate buffer. In some embodiments that may be combined with any of the previous embodiments, the excipient comprises glycerol. In some embodiments that may be combined with any of the previous embodiments, the excipient comprises a lipid carrier. In some embodiments that may be combined with any of the previous embodiments, the excipient comprises a nanoparticle carrier.

[0042] In some embodiments that may be combined with any of the preceding embodiments, the composition is a cosmetic composition. In some embodiments, the cosmetic composition is a skin care product.

[0043] Other aspects of the disclosure relate to a kit comprising any of the compositions described herein and instructions for administering the composition.

[0044] Other aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the level of one or more human collagens in a subject, the method comprising administering to the subject an effective amount of any of the compositions described herein.

[0045] Other aspects of the present disclosure relate to a method of enhancing, augmenting, strengthening and / or supplementing a subject's soft tissue, the method comprising administering to the subject an effective amount of any composition described herein. In some embodiments, the composition is injected into the subject's soft tissue.

[0046] Other aspects of the present disclosure relate to a method of improving the skin quality, condition and / or appearance of a subject in need thereof, the method comprising administering to the subject an effective amount of any composition described herein. In some embodiments, the condition is selected from sun damage, aging, UV exposure, rough texture, sagging skin, wrinkles, and any combination thereof.

[0047] Other aspects of the present disclosure relate to a method of reducing the appearance of one or more superficial depressions in the skin of a subject in need thereof, the method comprising applying to the subject an effective amount of any composition described herein. In some embodiments, the application of the composition continues for at least about three months, at least about six months, at least about nine months, or at least about 12 months to reduce the appearance of one or more superficial depressions in the subject's skin. In some embodiments, after applying the composition, the appearance of one or more superficial depressions in the subject's skin is reduced compared to the appearance of one or more superficial depressions in the subject's skin before applying the composition.

[0048] Other aspects of the present disclosure relate to a method of increasing and / or improving at least one of the texture, smoothness, elasticity or tension of a subject's skin in need, the method comprising applying an effective amount of any composition described herein to the subject. In some embodiments, after applying the composition, the subject's skin continues to maintain at least one of the texture, smoothness, elasticity or tension increase and / or improvement for at least about three months, at least about six months, at least about nine months, or at least about 12 months. In some embodiments, compared to the texture, smoothness, elasticity or tension of the subject's skin before applying the composition, after applying the composition, at least one of the texture, smoothness, elasticity or tension of the subject's skin is increased and / or improved.

[0049] In some embodiments that may be combined with any of the preceding embodiments, the subject's skin is aging skin. In some embodiments that may be combined with any of the preceding embodiments, the subject's skin has been damaged by exposure to ultraviolet light. In some embodiments that may be combined with any of the preceding embodiments, the subject's skin is wrinkled.

[0050] Other aspects of the present disclosure relate to a method of reducing one or more dermatological signs of aging in a subject in need thereof, the method comprising administering to the subject an effective amount of any composition described herein. In some embodiments, the reduction in one or more dermatological signs of aging is selected from: (a) treatment, reduction and / or prevention of fine lines and / or wrinkles; (b) reduction in skin pore size; (c) improvement in skin thickness, fullness and / or firmness; (d) improvement in skin smoothness, flexibility and / or softness; (e) improvement in skin tone, radiance and / or clarity; (f) improvement in procollagen and / or collagen production; (g) improvement in skin texture and / or promotion of retexturing; (h) improvement in skin (i) skin radiance and / or brightness is restored; (j) skin appearance that has diminished due to aging and / or menopause is improved; (k) skin moisturization is improved; (l) skin elasticity and / or resilience is increased; (m) skin sagging is treated, reduced and / or prevented; (n) skin firmness is improved; (o) pigment spots, mottled skin and / or acne scars are reduced; (p) skin optical properties in terms of light diffraction or reflection are improved; and (q) any combination thereof. In some embodiments, one or more dermatological signs of aging of the subject are reduced after applying the composition compared to one or more dermatological signs of aging of the subject before applying the composition.

[0051] In some embodiments that can be combined with any previous embodiment, the subject is a human. In some embodiments that can be combined with any previous embodiment, the composition is applied to the subject in a transdermal (systemic or local) manner, a transdermal manner, a subcutaneous manner and / or an intradermal manner. In some embodiments, the composition is applied by superficial injection. In some embodiments, the composition is applied to the subject in an intradermal manner. In some embodiments, the composition is applied to the subject once. In some embodiments, the composition is applied to the subject at least twice. In some embodiments, between applications, after at least about 15, at least about 30, at least about 60, at least about 90 or at least about 120 days. In some embodiments that can be combined with any previous embodiment, the composition is applied to one or more affected and / or unaffected areas of the subject. In some embodiments that can be combined with any previous embodiment, the skin is ground before application.

[0052] Other aspects of the present disclosure relate to a recombinant nucleic acid comprising a first polynucleotide encoding a first polypeptide, wherein the first polypeptide comprises a first human collagen, wherein the recombinant nucleic acid is a recombinant herpes simplex virus genome. In some embodiments, the recombinant nucleic acid comprises two or more copies of the first polynucleotide. In some embodiments, the recombinant herpes simplex virus genome is a recombinant HSV-1 genome, a recombinant HSV-2 genome, or any derivative thereof.

[0053] In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in a herpes simplex virus gene. In some embodiments, the herpes simplex virus gene is selected from the group consisting of infected cell protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41 and UL55. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in one or two copies of the ICP4 gene. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in the ICP22 gene. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in the UL41 gene. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in the ICPO gene. In some embodiments that can be combined with any previous embodiment, the recombinant herpes simplex virus genome includes an inactivation mutation in the ICP27 gene. In some embodiments that may be combined with any of the previous embodiments, the inactivating mutation is a deletion of the coding sequence of one or more genes.

[0054] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in the viral locus. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or two copies of the ICP4 viral locus. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in the ICP22 viral locus. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in the UL41 viral locus. In some embodiments that can be combined with any of the previous embodiments, HSV has reduced cytotoxicity compared to wild-type herpes simplex virus.

[0055] In some embodiments that may be combined with any of the preceding embodiments, the first human collagen is selected from the group consisting of COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6 In some embodiments that may be combined with any of the preceding embodiments, the first human collagen is selected from the group consisting of COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments that may be combined with any of the preceding embodiments, the nucleic acid sequence encoding the first human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments that may be combined with any of the preceding embodiments, the first human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments that may be combined with any of the preceding embodiments, the first human collagen is not COL7.

[0056] In some embodiments that may be combined with any of the preceding embodiments, the first polypeptide comprises: (a) a first human collagen; (b) another human collagen; and (c) a linker polypeptide that connects (a) to (b). In some embodiments, the linker polypeptide is a cleavable linker polypeptide. In some embodiments, the linker polypeptide comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 28-31. In some embodiments, the other human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the other human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments, the nucleic acid sequence encoding the other human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the other human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, the first human collagen and the other human collagen are different.

[0057] In some embodiments that may be combined with any of the preceding embodiments, the first polynucleotide encodes a polycistronic mRNA comprising: (a) a first open reading frame (ORF) encoding a first polypeptide; (b) a second ORF encoding an additional human collagen; and (c) an internal ribosome entry site (IRES) separating (a) and (b). In some embodiments, the nucleic acid sequence encoding the IRES has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleic acid sequence selected from SEQ ID NO: 22 or SEQ ID NO: 23. In some embodiments, the additional human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments, the nucleic acid sequence encoding the additional human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the additional human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, the first human collagen and the additional human collagen are different.

[0058] In some embodiments that may be combined with any of the previous embodiments, the recombinant nucleic acid further comprises a second polynucleotide encoding a second human collagen. In some embodiments, the recombinant nucleic acid comprises two or more copies of the second polynucleotide. In some embodiments, the second human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, and COL28. In some embodiments, the second human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7, and COL17. In some embodiments, the nucleic acid sequence encoding the second human collagen has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the second human collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, the first human collagen and the second human collagen are different.

[0059] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in the viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in one or two copies of the ICP4 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in the ICP22 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a second polynucleotide in the UL41 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or two copies of the ICP4 viral locus, and comprises a second polynucleotide in the ICP22 viral locus. In some embodiments, the recombinant herpes simplex virus genome comprises a first polynucleotide in one or two copies of the ICP4 viral locus, and comprises a second polynucleotide in the UL41 viral locus.

[0060] Other aspects of the present disclosure relate to a host cell comprising any of the recombinant nucleic acids described herein. In some embodiments, the host cell is a eukaryotic cell. In some embodiments, the host cell is a mammalian cell. In some embodiments, the host cell is a human cell or a non-human primate cell. In some embodiments, the host cell is a Vero cell. In some embodiments, the host cell is a complementary host cell.

[0061] Other aspects of the present disclosure relate to a method of collecting herpes simplex virus, the method comprising: (a) contacting a complementing host cell with any of the recombinant nucleic acids described herein; and (b) collecting the herpes simplex virus produced by the complementing host cell.

[0062] Other aspects of the present disclosure relate to a method of collecting herpes simplex virus, the method comprising: (a) culturing a host cell comprising any of the recombinant nucleic acids described herein; and (b) collecting the herpes simplex virus produced by the host cell. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] This patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0064] Figure 1A-Figure 1N Schematic diagram showing wild-type and modified herpes simplex virus genomes. Figure 1A The wild-type herpes simplex virus genome is shown. Figure 1BA modified herpes simplex virus genome showing a deletion comprising the coding sequences of ICP4 (two copies) and ICP22, having integrated at each of the ICP4 loci a polynucleotide containing the coding sequence of a first human collagen polypeptide operably linked to a heterologous promoter. Figure 1C Shown is a modified herpes simplex virus genome comprising a deletion of the coding sequence of ICP4 (two copies), having integrated at each of the ICP4 loci a polynucleotide containing the coding sequence of a first human collagen polypeptide operably linked to a heterologous promoter. Figure 1D A modified herpes simplex virus genome showing a deletion comprising the coding sequences of ICP4 (two copies) and ICP22, having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide containing 1) a coding sequence for a first human collagen polypeptide operably linked to a first heterologous promoter, and 2) a coding sequence for a second human collagen polypeptide operably linked to a second heterologous promoter. Both the first human collagen polypeptide and the second human collagen polypeptide are encoded on the same DNA strand. Figure 1E A modified herpes simplex virus genome showing a deletion comprising the coding sequence of ICP4 (two copies), having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide containing 1) a coding sequence for a first human collagen polypeptide operably linked to a first heterologous promoter, and 2) a coding sequence for a second human collagen polypeptide operably linked to a second heterologous promoter. Both the first human collagen polypeptide and the second human collagen polypeptide are encoded on the same DNA strand. Figure 1F A modified herpes simplex virus genome showing a deletion comprising coding sequences for ICP4 (two copies) and ICP22, having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide containing 1) a coding sequence for a first human collagen polypeptide operably linked to a first heterologous promoter, and 2) a coding sequence for a second human collagen polypeptide operably linked to a second heterologous promoter. The first human collagen polypeptide and the second human collagen polypeptide are encoded on opposite DNA strands. Figure 1G A modified herpes simplex virus genome showing a deletion comprising a coding sequence for ICP4 (two copies), having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide containing 1) a coding sequence for a first human collagen polypeptide operably linked to a first heterologous promoter, and 2) a coding sequence for a second human collagen polypeptide operably linked to a second heterologous promoter. The first human collagen polypeptide and the second human collagen polypeptide are encoded on opposite DNA strands. Figure 1HA modified herpes simplex virus genome showing a deletion comprising the coding sequences of ICP4 (two copies) and ICP22, having a polynucleotide integrated at each of the ICP4 loci encoding a polycistronic mRNA operably linked to a heterologous promoter. The polycistronic mRNA contains the coding sequence of a first human collagen polypeptide and the coding sequence of a second human collagen polypeptide separated by an internal ribosome entry site (IRES). Fig. 1I A modified herpes simplex virus genome showing a deletion comprising the coding sequence of ICP4 (two copies), having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide encoding a polycistronic mRNA operably linked to a heterologous promoter. The polycistronic mRNA contains the coding sequence of a first human collagen polypeptide and the coding sequence of a second human collagen polypeptide separated by an internal ribosome entry site (IRES). Figure 1J A modified herpes simplex virus genome showing a deletion comprising the coding sequences of ICP4 (two copies) and ICP22, having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide containing the coding sequence of a chimeric polypeptide operably linked to a heterologous promoter. The chimeric polypeptide comprises the amino acid sequence of a first human collagen polypeptide and the amino acid sequence of a second human collagen polypeptide separated by a cleavable linker. Figure 1K A modified herpes simplex virus genome showing a deletion comprising the coding sequence of ICP4 (two copies), having a polynucleotide integrated at each of the ICP4 loci, the polynucleotide containing the coding sequence of a chimeric polypeptide operably linked to a heterologous promoter. The chimeric polypeptide comprises the amino acid sequence of a first human collagen polypeptide and the amino acid sequence of a second human collagen polypeptide separated by a cleavable linker. Figure 1L A modified herpes simplex virus genome showing a deletion comprising coding sequences of ICP4 (two copies) and ICP22, having a first polynucleotide integrated at each of the ICP4 loci and a second polynucleotide integrated at the ICP22 locus, wherein the first polynucleotide contains the coding sequence of a first human collagen polypeptide operably linked to a heterologous promoter, and the second polynucleotide contains the coding sequence of a second human collagen polypeptide operably linked to a heterologous promoter. Figure 1M A modified herpes simplex virus genome showing a deletion comprising the coding sequences of ICP4 (two copies), ICP22, and UL41, having a first polynucleotide integrated at each of the ICP4 loci and a second polynucleotide integrated at the UL41 locus, wherein the first polynucleotide contains the coding sequence of a first human collagen polypeptide operably linked to a heterologous promoter, and the second polynucleotide contains the coding sequence of a second human collagen polypeptide operably linked to a heterologous promoter. Figure 1NA modified herpes simplex virus genome showing a deletion comprising the coding sequences of ICP4 (two copies) and UL41, having a first polynucleotide integrated at each of the ICP4 loci and a second polynucleotide integrated at the UL41 locus, wherein the first polynucleotide contains the coding sequence of a first human collagen polypeptide operably linked to a heterologous promoter, and the second polynucleotide contains the coding sequence of a second human collagen polypeptide operably linked to a heterologous promoter.

[0065] Figure 2A-2B Schematic diagram showing a replication-defective herpes simplex type 1 virus carrying a human collagen 7 (COL7) expression cassette. Figure 2A A schematic diagram showing the virus "KCA211". Figure 2B A schematic diagram showing the virus "SAR-COL7".

[0066] Figure 3A-3B Shown is the expression of human COL7 in HaCaT cells infected with KCA211 or SAR-COL7 at the indicated MOIs. Figure 3A Human COL7 expression in HaCaT cells infected with KCA211 or SAR-COL7 at the indicated MOIs is shown as assessed by qPCR. Data are shown as fold change relative to SAR-COL7 after normalization to GAPDH. Figure 3B Shown is human COL7 expression in uninfected HaCaT cells or HaCaT cells infected with KCA211 or SAR-COL7 at the indicated MOIs, as assessed by Western blot analysis.

[0067] Figure 4A-4B Immunofluorescence images showing human COL7 expression in mock-infected primary human cells isolated from healthy patients (Normal) and mock- or SAR-COL7-infected primary human cells isolated from patients with recessive dystrophic epidermolysis bullosa (RDEB). Figure 4A Shown is human COL7 expression in wild-type and RDEB primary human keratinocytes mock-infected or RDEB primary human keratinocytes infected with SAR-COL7 at the indicated multiplicity of infection (MOI). Figure 4B Shown is human COL7 expression in wild-type and RDEB primary human fibroblasts mock-infected or RDEB primary human fibroblasts infected with SAR-COL7 at the indicated MOIs.

[0068] Figure 5A-Figure 5B Shown are quantitative PCR analyses of human COL7 expression in mock-infected primary human cells isolated from healthy patients and mock- or SAR-COL7-infected primary human cells isolated from patients with recessive dystrophic epidermolysis bullosa (EB). Figure 5A Human COL7 expression is shown in mock-infected wild-type (N-HDK) and RDEB (EB-HDK) primary human keratinocytes or in RDEB primary human keratinocytes infected with SAR-COL7 at the indicated MOIs. COL7 expression is shown as relative fold change compared to mock-infected wild-type primary human keratinocytes. Figure 5B Human COL7 expression is shown in mock-infected wild-type (N-HDF) and RDEB (EB-HDF) primary human fibroblasts or in RDEB primary human fibroblasts infected with SAR-COL7 at the indicated MOIs. COL7 expression is shown as relative fold change compared to mock-infected wild-type primary human fibroblasts.

[0069] Figure 6A-6B Cell adhesion of uninfected (control) or SAR-COL7 infected RDEB primary human keratinocytes to untreated (plastic) or treated wells of a microplate is shown. Fig. 6A Cell adhesion to untreated wells (plastic) or wells treated with increasing concentrations of rat tail collagen 1 is shown. Figure 6B Cell adhesion to untreated wells (plastic) or wells treated with increasing concentrations of human plasma fibronectin is shown.

[0070] Figure 7 Representative immunofluorescence images of human COL7 expression and deposition at the basement membrane zone (BMZ) at day 5 in organotypic cultures established with RDEB primary human keratinocytes and fibroblasts infected with SAR-COL7 are shown. Both keratinocytes and fibroblasts were infected in situ at the indicated MOIs after culture establishment.

[0071] Figure 8A-8D Shown are human COL7A1 transcript and genomic levels observed in uninfected mouse skin (control) or in mouse skin following topical or intradermal delivery of SAR-COL7, as assessed by qPCR. Error bars represent SEM. Fig. 8A Shown are human COL7A1 transcript levels per 100 ng of total RNA in mouse skin at day 3 after infection. Figure 8B Shown are the human COL7A1 DNA copies / 100 ng total DNA in mouse skin at day 3 after infection. Figure 8C Shown are human COL7A1 transcript levels per 100 ng of total RNA in mouse skin at day 6 after infection. Fig.8D Shown are the human COL7A1 DNA copies / 100 ng total DNA in mouse skin at day 6 after infection.

[0072] Figure 9A-9BRepresentative immunofluorescence images showing human COL7 expression in mouse skin after delivery of SAR-COL7. Fig.9A Representative immunofluorescence images showing human COL7 expression in mouse skin following intradermal delivery of SAR-COL7. Fig. 9B Representative immunofluorescence images showing human COL7 expression in mouse skin after topical delivery of SAR-COL7.

[0073] Figure 10A-10B Shown are human COL7A1 transcript and genomic levels observed in BALB / c mouse skin following intradermal delivery of vehicle, SAR-COL7, or KCA211, as assessed by qPCR. Fig. 10A Shown are human COL7A1 transcript levels per 100 ng total RNA in BALB / c mouse skin. Fig. 10B Shown are the human COL7A1 DNA copy numbers per 100 ng total DNA in BALB / c mouse skin.

[0074] Figure 11A-11B Shown are human COL7A1 transcript and genome levels observed at each injection site in hypoeffective mouse skin following high-dose intradermal delivery of HSV-GFP (GFP control) or SAR-COL7, as assessed by qPCR. Each bar represents a single sample at the indicated time point. Fig.11A Human COL7A1 transcript levels per 100 ng total RNA in hypoperforming mouse skin are shown. Fig. 11B The human COL7A1 DNA copy number / 100 ng total DNA in hypoperfective mouse skin is shown.

[0075] Figure 12A-12B Representative immunofluorescence images showing human COL7 expression in hypofunctional mouse skin following high-dose intradermal delivery of HSV-GFP (GFP control) or SAR-COL7. Fig. 12A Shown are control (GFP) and SAR-COL7 immunofluorescence images obtained from hypoeffective mouse 1 (collected on day 3) at 10X and 20X magnification. Fig. 12B Shown are SAR-COL7 immunofluorescence images obtained from subeffective mice 2 and 3 (collected on day 7). The graph represents a tiled image of 16 fields acquired with a 10x lens, capturing the entire skin cross section.

[0076] Fig.13 Shown are H&E stained samples from subeffective mice 1, 2, and 3 (collected on days 3 and 4). Samples were taken from untreated subeffective mouse skin and subeffective mouse skin after intradermal delivery of HSV-GFP or SAR-COL7.

[0077] Figure 14A-14B Representative electron micrograph images showing human COL7 expression in hypomorphic mouse skin following intradermal delivery of SAR-COL7. The stratum compactum is the indicated dark band through the center of the image; black dots are stained NC domains of human COL71, and blue arrows indicate the formation of anchoring fibrils. Fig.14A Electron micrograph images of infected hypoplastic mouse skin stained with an antibody specific for the NC2 domain of human COL7 (LH24) are shown. Fig. 14B Electron micrograph images of infected hypoplastic mouse skin stained with an antibody specific for the NC1 domain of human COL7 (NP185) are shown.

[0078] Figure 15A-15B Shown are human COL7A1 transcript and genomic levels observed at each injection site in hypoperforming mouse skin following low-dose intradermal delivery of SAR-COL7, as assessed by qPCR. Each bar represents a single sample at the indicated time point. Fig.15A Human COL7A1 transcript levels per 100 ng total RNA in hypoperforming mouse skin are shown. Fig. 15B The human COL 7A1 DNA copy number / 100 ng total DNA in the skin of hypoplastic mice is shown.

[0079] Fig.16 Representative immunofluorescence images showing human COL7 expression in hypodermal mouse skin (from mouse 1) following low-dose intradermal delivery of SAR-COL7.

[0080] Figure 17A-17C Shown are human COL1A1 and COL1A2 nucleic acid and protein analyses in the context of Vero cells infected with the indicated clones of HSV encoding COL1A1 alone (inserted into the ICP4 locus) or COL1A1 and COL1A2 (inserted into the ICP4 and ICP22 loci, respectively). Fig.17A Shown are the levels of human COL1A1 transcripts present in Vero cells 5 days after infection with the indicated HSV clones, as determined by qRT-PCR analysis. Data from two replicates ± SEM are presented. Fig. 17B Shown are the levels of human COL1A2 transcripts present in Vero cells 5 days after infection with the indicated HSV clones, as determined by qRT-PCR analysis. Data from two replicates ± SEM are presented. Fig. 17CWestern blot analysis showing human COL1A1 and COL1A2 protein expression in Vero cells 5 days after infection with the indicated COL1A1 / COL1A2 positive clones as determined by qRT-PCR. Uninfected (mock) Vero cells were used as negative control; GAPDH was used as loading control.

[0081] Fig.18 Western blot analysis showing human COL1A1 and COL1A2 protein expression in Vero cells 5 days after infection with an HSV isolate encoding a COL1A1-IRES-COL1A2 sequence inserted into the ICP4 locus (IRES-Isolate 6). Infection with an isolate without an IRES construct (no insert) was used as a negative control; GAPDH was used as a loading control.

[0082] Figure 19A-19B Shown are human COL3 nucleic acid and protein analysis in the context of immortalized human keratinocytes (HaCaTs) infected with C3vec01. Fig.19A Shown are the levels of human COL3A1 transcripts present in immortalized human keratinocytes (HK) after infection with C3vec01 at the indicated MOIs. Uninfected (mock) and HSV-mCherry infected (mCherry) cells were used as negative controls. Data ± SEM of two replicates are presented. Fig.19B Representative immunofluorescence images showing human COL3 protein expression in immortalized human keratinocytes 48 hours after infection with C3vec01 at the indicated MOIs. Uninfected (mock) cells were used as negative controls.

[0083] Figure 20A-20B Shown are human COL3 nucleic acid and protein analysis in the context of immortalized human dermal fibroblasts (HDFs) infected with C3vec01. Fig. 20A Shown are the levels of human COL3A1 transcripts present in immortalized human dermal fibroblasts (HDFs) following infection with C3vec01 at the indicated MOIs. Uninfected (mock) and HSV-mCherry infected (mCherry) cells were used as negative controls. Data from two replicates ± SEM are presented. Fig. 20B Representative immunofluorescence images showing human COL3 protein expression in immortalized human dermal fibroblasts 48 hours after infection with C3vec01 at the indicated MOIs. Uninfected (mock) cells were used as negative controls.

[0084] Figure 21A-Figure 21DShown are human COL3 nucleic acid and protein analysis in the context of aged primary human fibroblasts (HDFs) derived from two different suppliers infected with C3vec01 at the indicated MOIs. Fig.21A Shown are the levels of human COL3A1 transcripts present in primary HDFs (Vendor 1) collected from a 65-year-old female patient or a 73-year-old male patient following infection with C3vec01 at the indicated MOIs. Uninfected (mock) cells were used as negative controls. Data from two replicates ± SEM are presented. Fig. 21B Western blot analysis showing human COL3A1 protein expression in primary HDFs collected from a 73-year-old male patient (Supplier 1) following infection with C3vec01 at the indicated MOIs. Uninfected (mock) cells were used as negative controls; recombinant human COL3A1 (rCOL3A1) was used as a positive control; GAPDH was used as a loading control. Fig. 21C Shown are the levels of human COL3A1 transcripts present in primary HDFs (Vendor 2) collected from a 75-year-old female patient or a 73-year-old male patient following infection with C3vec01 at the indicated MOIs. Uninfected (mock) cells were used as negative controls. Data from two replicates ± SEM are presented. Fig.21D Western blot analysis showing human COL3A1 protein expression in primary HDFs collected from a 75-year-old female patient (Supplier 2) following infection with C3vec01 at the indicated MOIs. Uninfected (mock) cells were used as negative controls; recombinant human COL3A1 (rCOL3A1) was used as a positive control; GAPDH was used as a loading control.

[0085] Figure 22A-22B Shown are human COL3 nucleic acid and protein analysis in the context of immortalized human dermal fibroblasts (HDFs) after UV exposure. Fig.22A Shown are the concentrations of COL3 secreted into the supernatant of cultured HDFs 24 hours after exposure to various doses and times of UV light, as assessed by ELISA. Supernatant collected from non-UV exposed (-UV) HDFs cultured in parallel was used as a control. Fig. 22B Shown are the levels of human COL3A1 transcripts present in UV-exposed immortalized human dermal fibroblasts (HDFs) following infection with C3vec01 at the indicated MOIs. Uninfected (mock) and HSV-mCherry infected (mCherry) cells were used as negative controls. Data ± SEM of two replicates are presented.

[0086] Figure 23A-23CShown are COL3 nucleic acid and protein analyses of skin biopsies taken from young (6-8 weeks old) and old (approximately 13 months old) C57BL / 6 mice treated with control or C3vec01 48 hours after intradermal application. Fig.23A Shown are the levels of human COL3A 1 DNA present in skin biopsies taken from young and old mice 48 hours after intradermal administration of C3vec01 or vehicle control, as assessed by qPCR analysis. Fig. 23B Shown are the levels of human COL3A1 transcripts present in skin biopsies taken from young and old mice 48 hours after intradermal administration of C3vec01 or vehicle control, as assessed by qRT-PCR analysis. For each condition in qPCR and qRT-PCR analysis, data are presented as the mean ± SEM of four tissue samples (two replicates / tissue sample). Fig.23C Representative immunofluorescence images of human COL3 expression in skin biopsies taken from young and old mice 48 hours after intradermal administration of C3vec01 are shown. Young mice administered vehicle alone intradermally served as negative controls. DAPI staining was used to visualize nuclei.

[0087] Figure 24A-24B Shown is the expression of wild-type (WT) human LamB3 in Vero cells infected with the indicated viral isolates. Fig.24A Shown is the expression of wild-type human LAMB3 in infected Vero cells, as assessed by qPCR analysis. Fig. 24B Expression of wild-type human LamB3 protein in infected Vero cells, as assessed by Western blotting, is shown.

[0088] Fig.25 Expression of wild-type (WT) or codon-optimized (CO) human LamB3 protein in Vero cells infected with the indicated virus isolates, as assessed by Western blot, is shown. Uninfected Vero cells were used as negative control.

[0089] Fig.26 Shown is the expression of wild-type (WT) or codon-optimized (CO) human LamB3 protein in primary human keratinocytes infected with the indicated viral isolates, as assessed by Western blot. Uninfected primary keratinocytes served as negative control.

[0090] Figure 27A-27C Shown is the expression of wild-type (WT) and codon-optimized (CO) human LamC2 in Vero cells infected with the indicated viral isolates. Fig.27A Shown is the expression of wild-type human LAMC2 in infected Vero cells, as assessed by qPCR analysis. Fig.27BShown is the expression of codon-optimized human LAMC2 in infected Vero cells, as assessed by qPCR analysis. Fig.27C Expression of wild-type and codon-optimized human LamC2 proteins in infected Vero cells is shown, as assessed by Western blot. The boxed virus isolate "LGA" expressing codon-optimized LamC2 was selected for additional experiments.

[0091] Figure 28A-28C Shown is human LAMC2 expressed from the viral isolate "LGA" in immortalized primary human keratinocytes infected at the indicated multiplicity of infection (MOI). Fig.28A Shown are viral genome copy numbers in primary immortalized human keratinocytes following infection with viral isolate "LGA" at the indicated MOIs. Fig.28B Shown are transcript levels of codon-optimized LAMC2 expressed in primary immortalized human keratinocytes following infection with the viral isolate "LGA" at the indicated MOIs. Fig.28C Shown is the expression of human LamC2 protein in primary immortalized human keratinocytes following infection with the viral isolate "LGA" at the indicated MOIs, as assessed by Western blot.

[0092] Figure 29A-Figure 29D Shown are LAMC2 nucleic acid and protein analysis of skin biopsies taken from mice treated with control (vehicle) or HSV isolate "LGA" 72 hours after intradermal application. Fig.29A Schematic diagram showing the intradermal injection sites on treated animals.

[0093] Fig.29B Shown are the levels of human LAMC2 DNA present in skin biopsies taken from mice 72 hours following intradermal administration of HSV isolate LGA or vehicle control, as assessed by qPCR analysis. Fig.29C Shown are the levels of human LAMC2 transcripts present in skin biopsies taken from mice 72 hours after intradermal administration of HSV isolate LGA or vehicle control, as assessed by qRT-PCR analysis. For each condition in qPCR and qRT-PCR analysis, data are presented as the mean ± SEM of two replicates. Fig.29D Representative immunofluorescence images of human LAMC2 expression in skin biopsies taken from mice 72 hours after intradermal administration of HSV isolate LGA are shown. Sites where vehicle alone was administered intradermally served as negative controls. DAPI staining was used to visualize nuclei; pKal staining was used to visualize mouse laminin-332. DETAILED DESCRIPTION

[0094] In some embodiments, the present disclosure relates to recombinant nucleic acids (e.g., recombinant herpes virus genomes) encoding one or more beauty proteins, and the use of these recombinant nucleic acids in viruses (e.g., herpes viruses), compositions, preparations, medicaments, and / or methods for delivering one or more beauty proteins to the skin, such as onto the skin, into the skin, and / or through the skin (e.g., to the dermal ECM). In some embodiments, the present disclosure relates to recombinant nucleic acids (e.g., recombinant herpes virus genomes) encoding one or more beauty proteins, and the use of these recombinant nucleic acids in viruses (e.g., herpes viruses), compositions, preparations, medicaments, and / or methods for increasing, strengthening, and / or supplementing one or more dermal ECM proteins (e.g., one or more collagens). In some embodiments, the present disclosure relates to recombinant nucleic acids (e.g., recombinant herpes virus genomes) encoding one or more beauty proteins, and the use of these recombinant nucleic acids in viruses (e.g., herpes viruses), compositions, preparations, medicaments, and / or methods in cosmetic situations (e.g., to reduce one or more dermatological signs of aging). In some embodiments, the disclosure relates to a composition comprising a recombinant herpesvirus vector and includes a method of delivering the recombinant herpesvirus vector to the skin of a mammal, to the skin of a mammal and / or through the skin of a mammal, wherein the recombinant herpesvirus vector is included in a promoter operable in a mammalian cell and is expressed to realize a heterologous nucleic acid for a cosmetic effect in the skin of a mammal. Heterologous nucleic acid can be delivered to a mammalian target skin cell, including making the epidermis, dermis or subcutaneous tissue of the mammal and a composition comprising a recombinant herpesvirus vector on the epidermis, dermis or subcutaneous tissue, to the epidermis, dermis or subcutaneous tissue, and / or transporting the recombinant herpesvirus vector through the epidermis, dermis or subcutaneous tissue, and introducing it to contact under the conditions relied upon in the target skin cell expressed therein. Without wishing to be bound by theory, it is believed that applying one or more of the recombinant nucleic acids, viruses and / or preparations described herein to an individual will allow the production of functional dermal ECM proteins (such as human collagen) in the individual to increase. In addition, without wishing to be bound by theory, it is believed that increasing, strengthening and / or supplementing the level of cosmetic proteins in an individual by administering one or more of the recombinant nucleic acids, viruses and / or formulations described herein will result in at least one of the following: 1) enhancement, strengthening and / or supplementation of soft tissue; 2) improvement in skin quality, condition and / or appearance; 3) reduction in one or more superficial depressions (e.g., wrinkles) in the skin; 4) improvement in the texture, smoothness, elasticity and / or tension of the skin; and / or 5) alleviation of one or more dermatological signs of aging. Finally, without wishing to be bound by theory, it is believed that the recombinant nucleic acids, viruses, compositions and methods described herein provide a novel strategy for delivering functional cosmetic proteins in a cosmetic setting.

[0095] The following description sets forth exemplary methods, parameters, etc. However, it should be appreciated that such description is not intended as a limitation on the scope of the present disclosure, but is instead provided as a description of exemplary embodiments.

[0096] I. General Technology

[0097] The techniques and procedures described or referenced herein are generally well understood by those skilled in the art and are routinely employed by those skilled in the art using routine methods, such as the widely used methods described in Sambrook et al., Molecular Cloning.·A Laboratory Manual 3rd Edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (FM Ausubel et al., eds., (2003)); the book Methods in Enzymology (Academic Press, Inc.); PCR 2: A Practical Approach (MJ MacPherson, BD Hames and GR Taylor, eds. (1995)); Harlow and Lane, eds. (1988); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (JE Cellis, ed., 1998) Academic Press; Animal ... Culture (RI Freshney), 1987); Introduction to Cell and Tissue Culture (JPMather and PERoberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A.Doyle, JBGriffiths and DG Newell, 1993-8) J.Wiley and Sons; Gene Transfer Vectors for Mammalian Cells (JMMiller and MPCalos, 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., 1994); Short Protocols in Molecular Biology (Wiley and Sons, 1999).

[0098] II. Definitions

[0099] Before describing the present disclosure in detail, it is to be understood that the present disclosure is not limited to particular compositions or biological systems, which may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0100] As used herein, the singular forms "a", "an", and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a molecule" optionally includes a combination of two or more of those molecules, and so on.

[0101] As used herein, the term "and / or" may include any and all combinations of one or more of the associated listed items. For example, the term "a and / or b" may refer to "a alone", "b alone", "a or b", or "a and b"; the term "a, b, and / or c" may refer to "a alone", "b alone", "c alone", "a or b", "a or c", "b or c", "a, b or c", "a and b", "a and c", "b and c", or "a, b and c"; etc.

[0102] As used herein, the term "about" refers to the usual error range for the corresponding value that is readily known to those skilled in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments related to that value or parameter itself.

[0103] It should be understood that aspects and embodiments of the present disclosure include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0104] As used herein, the terms "polynucleotide", "nucleic acid sequence", "nucleic acid" and variations thereof will be generic to polydeoxyribonucleotides (containing 2-deoxy-D-ribose), polyribonucleotides (containing D-ribose), any other type of polynucleotide that is an N-glycoside of a purine or pyrimidine base, and other polymers containing non-nucleotide backbones, provided that the polymer contains nucleoside bases in a configuration that permits base pairing and base stacking as seen in DNA and RNA. Thus, these terms include known types of nucleic acid sequence modifications, such as substitution of one or more naturally occurring nucleotides with an analog, as well as internucleotide modifications.

[0105] As used herein, a nucleic acid is "operably linked" or "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it positions the site so as to promote translation. Generally, "operably linked" or "operably linked" means that the DNA sequences being linked are contiguous.

[0106] As used herein, the term "vector" refers to a discrete element used to introduce a heterologous nucleic acid into a cell to achieve its expression or replication. Expression vectors include vectors capable of expressing a nucleic acid operably linked to a regulatory sequence, such as a promoter region, which is capable of achieving expression of the nucleic acid. Thus, an expression vector may refer to a DNA or RNA construct, such as a plasmid, a phage, a recombinant virus or other vector, that results in expression of a nucleic acid upon introduction into an appropriate host cell. Suitable expression vectors are well known to those skilled in the art and include those that can replicate in eukaryotic cells, as well as those that remain free or are integrated into the host cell genome.

[0107] As used herein, "open reading frame" or "ORF" refers to a continuous stretch of DNA or RNA nucleic acid encoding a protein or polypeptide. Typically, the nucleic acid comprises a translation initiation signal or start codon such as ATG or AUG, and a stop codon.

[0108] As used herein, "untranslated region" or "UTR" refers to non-translated nucleic acid at the 5' and / or 3' end of an open reading frame. Including one or more UTRs in a polynucleotide can affect post-transcriptional regulation, mRNA stability and / or translation of the polynucleotide.

[0109] As used herein, the term "transgenic" refers to a polynucleotide that can be transcribed into RNA and translated and / or expressed under appropriate conditions after being introduced into a cell. In some aspects, it confers desired properties to the cell into which it is introduced, or otherwise results in a desired cosmetic, therapeutic or diagnostic effect.

[0110] As used herein, the terms "polypeptide," "protein," and "peptide" are used interchangeably and may refer to a polymer of two or more amino acids.

[0111] As used herein, "subject," "host," or "individual" refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sport, or pet animals, such as dogs, horses, cats, cows, and animals used in research, such as mice, rats, hamsters, rabbits, and non-human primates, etc. In some embodiments, the mammal is a human.

[0112] As used herein, the term "pharmaceutical formulation" or "pharmaceutical composition" refers to a preparation that is in a form that permits the biological activity of one or more active ingredients to be effective, and does not contain additional components that are unacceptably toxic to the subject to which the composition or formulation will be administered. "Pharmaceutically acceptable" excipients (e.g., vehicles, additives) are those that can reasonably be administered to a subject to provide an effective dose of the active ingredient or ingredients employed.

[0113] As used herein, "transdermal administration" or "administered transdermally" refers to delivering a composition to a subject by contacting a formulation containing the composition directly or otherwise with all ("systemic") or a portion ("topical") of the subject's skin. The term encompasses several routes of administration, including, but not limited to, topical and transdermal. Topical administration can be used as a means to deliver a composition to the epidermis or dermis of a subject, or a specific layer thereof.

[0114] As used herein, "treatment" refers to clinical intervention intended to alter the natural processes of the individual or cell being treated during the course of a clinical condition. Desirable therapeutic effects include reducing the rate of progression of a disease / disorder / defect, ameliorating or alleviating the disease / disorder / defect state, and alleviating or improving prognosis. For example, an individual is successfully "treated" if one or more symptoms associated with dermatological aging are reduced, alleviated, or eliminated, including reduction or elimination of wrinkles.

[0115] As used herein, the term "delaying the progression of a disease / disorder / defect" means to postpone, hinder, slow, slow, stabilize, and / or delay the development of the disease / disorder / defect (e.g., skin wrinkles). This delay may be of varying lengths or times, depending on the history of the disease / disorder / defect and / or the individual being treated. As will be apparent to one of ordinary skill in the art, a substantial or significant delay may actually encompass prevention, in that the individual does not manifest the disease / disorder / defect.

[0116] III. Recombinant Nucleic Acids

[0117] Certain aspects of the present disclosure relate to recombinant nucleic acids (e.g., isolated recombinant nucleic acids) comprising one or more polynucleotides encoding cosmetic proteins (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.). Any suitable cosmetic protein described herein or known in the art may be encoded by the polynucleotides of the present disclosure, including, for example, collagen, fibronectin, elastin, aphrodisiac, vitronectin / vitronectin receptor, laminin, neuromodulators, fibrillin, additional dermal ECM proteins, and the like. In some embodiments, the cosmetic protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the cosmetic protein is collagen, elastin, fibronectin, or laminin (e.g., human collagen, elastin, fibronectin, or laminin).

[0118] In some embodiments, the present disclosure relates to a recombinant nucleic acid (e.g., an isolated recombinant nucleic acid) comprising one or more polynucleotides encoding collagen (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.). In some embodiments, the collagen is human collagen. In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides encoding homotrimeric collagen (e.g., homotrimeric human collagen, such as human collagen 3 (e.g., including three COL3A1 (COL3) polypeptides) or human collagen 7 (e.g., including three COL7A1 (COL7) polypeptides)). In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides encoding heterotrimeric collagen (e.g., heterotrimeric human collagen, such as human collagen 1 (e.g., including two COL1A1 (COL1-1) polypeptides and one COL1A2 (COL1-2) polypeptide) or human collagen 4 (e.g., including two COL4A1 (COL4-1) polypeptides and one COL4A2 (COL4-2) polypeptide)). In some embodiments, the present disclosure relates to recombinant nucleic acids comprising one or more polynucleotides encoding homotrimeric collagens and heterotrimeric collagens (e.g., recombinant nucleic acids comprising one or more polynucleotides encoding human collagen 1 and human collagen 3). In some embodiments, the present disclosure relates to recombinant nucleic acids comprising one or more polynucleotides encoding human collagen 1. In some embodiments, the present disclosure relates to recombinant nucleic acids comprising one or more polynucleotides encoding human collagen 3.

[0119] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising a first polynucleotide encoding a first polypeptide, the first polypeptide comprising a first beauty protein (e.g., a first human collagen). In some embodiments, the first polypeptide consists essentially of or consists of a first beauty protein (e.g., essentially of or consists of a first human collagen). In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising a first polynucleotide encoding a first polypeptide, the first polypeptide comprising: a first beauty protein (e.g., a first human collagen), a linker polypeptide, and another beauty protein (e.g., another human collagen). In some embodiments, the first beauty protein and another beauty protein (e.g., a first human collagen and another human collagen) are the same. In some embodiments, the first beauty protein and another beauty protein (e.g., a first human collagen and another human collagen) are different. In some embodiments, the linker polypeptide is a cleavable linker polypeptide.

[0120] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising a first polynucleotide encoding a first polypeptide, the first polypeptide comprising a first cosmetic protein (e.g., a first human collagen), wherein the first polynucleotide encodes a polycistronic mRNA, the polycistronic mRNA comprising: a first open reading frame (ORF) encoding the first polypeptide, an internal ribosome entry site (IRES), and a second ORF encoding an additional cosmetic protein (e.g., an additional human collagen). In some embodiments, the first cosmetic protein and the additional cosmetic protein (e.g., the first human collagen and the additional human collagen) are the same. In some embodiments, the first cosmetic protein and the additional cosmetic protein (e.g., the first human collagen and the additional human collagen) are different.

[0121] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising a first polynucleotide encoding a first polypeptide, the first polypeptide comprising a first cosmetic protein (e.g., a first human collagen); and a second polynucleotide encoding a second cosmetic protein (e.g., a second human collagen). In some embodiments, the first cosmetic protein and the second cosmetic protein (e.g., a first human collagen and a second human collagen) are the same. In some embodiments, the first cosmetic protein and the second cosmetic protein (e.g., a first human collagen and a second human collagen) are different.

[0122] In some embodiments, the recombinant nucleic acid is a vector. In some embodiments, the recombinant nucleic acid is a viral vector. In some embodiments, the recombinant nucleic acid is a herpes virus vector. In some embodiments, the recombinant nucleic acid is a herpes simplex virus amplicon. In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus genome. In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome.

[0123] Polynucleotides encoding beauty proteins

[0124] Polynucleotide encoding collagen

[0125] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of a collagen gene. The coding sequence of any collagen gene (including any subtype thereof) from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, human collagen genes (see, for example, NCBI gene identifiers: 1277, 1278, 1281, 1282, 1284, 1291, 1294, 1308, etc.), mouse collagen genes (see, for example, NCBI gene identifiers: 12842, 12843, 12825, 12826, 12827, 12833, 12836, 12821, etc.), chimpanzee collagen genes (see, for example, NCBI gene identifiers: 104001053, 45511 7, 459815, 452689, 452661, 450204, 101056895, 101058306, etc.), rat collagen gene (see, for example, NCBI gene identifiers: 29393, 84352, 84032, 290905, 306628, 294337, 301012, 294027, etc.), rabbit collagen gene (see, for example, NCBI gene identifiers: 100347598, 100008997, 100009177, 100358256, 100358522, 100343947, 100356561, 100339335, etc.), etc. Methods for identifying collagen gene homologs / orthologs from additional species are known to those of ordinary skill in the art and include, for example, the use of nucleic acid sequence alignment programs such as blasm kit. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any collagen gene (and / or its coding sequence) described herein or known in the art.

[0126] In some embodiments, the polynucleotides of the present disclosure include codon optimized variants of the coding sequences of any collagen genes described herein or known in the art. In some embodiments, the use of codon optimized variants of the coding sequences of collagen genes increases the stability and / or yield of heterologous expression of the encoded collagen in target cells (such as cells of the epidermis and / or dermis) compared to the stability and / or yield of heterologous expression of the corresponding non-codon optimized wild-type sequence. Any suitable method for codon optimization of sequences for expression in one or more target cells (e.g., one or more human cells) known in the art can be used, including, for example, by the method described by Fath et al. (PLoS One. 2011 March 3; 6 (3): e17596).

[0127] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising a coding sequence of a human collagen gene. Any suitable human collagen gene known in the art (including any subtype thereof) can be encoded by the nucleic acids of the present disclosure, including, for example, the COL1A1 gene (see, e.g., NCBI gene identifier: 1277; SEQ ID NO: 1), the COL1A2 gene (see, e.g., NCBI gene identifier: 1278; SEQ ID NO: 3), the COL2A1 gene (see, e.g., NCBI gene identifier: 1280), the COL3A1 gene (see, e.g., NCBI gene identifier: 1281; SEQ ID NO: 5), the COL4A1 gene (see, e.g., NCBI gene identifier: 1282; SEQ ID NO: 6), the COL5A1 gene (see, e.g., NCBI gene identifier: 1283; SEQ ID NO: 7), the COL6A1 gene (see, e.g., NCBI gene identifier: 1284; SEQ ID NO: 8), the COL7A1 gene (see, e.g., NCBI gene identifier: 1285; SEQ ID NO: 9), the COL8A1 gene (see, e.g., NCBI gene identifier: 1286; SEQ ID NO: 10), the COL9A1 gene (see, e.g., NCBI gene identifier: 1287; SEQ ID NO: 11), the COL1A2 gene (see, e.g., NCBI gene identifier: 1288; SEQ ID NO: 12), the COL1A2 gene (see, e.g., NCBI gene identifier: 1289; SEQ ID NO: 13), the COL2A1 gene (see, e.g NO:7), COL4A2 gene (see, e.g., NCBI gene identifier: 1284), COL4A3 gene (see, e.g., NCBI gene identifier: 1285), COL4A4 gene (see, e.g., NCBI gene identifier: 1286), COL4A5 gene (see, e.g., NCBI gene identifier: 1287), COL4A6 gene (see, e.g., NCBI gene identifier: 1288), COL5A1 gene (see, e.g., NCBI gene identifier: 1289), COL5A2 gene (see, e.g., NCBI gene identifier: 1290), COL5A3 gene (see, e.g., NCBI gene identifier: 50509), COL6A1 gene (see, e.g., NCBI gene identifier: 1291; SEQ ID NO:7), COL4A2 gene (see, e.g., NCBI gene identifier: 1284), COL4A3 gene (see, e.g., NCBI gene identifier: 1285), COL4A4 gene (see, e.g., NCBI gene identifier: 1286), COL4A5 gene (see, e.g., NCBI gene identifier: 1287), COL4A6 gene (see, e.g., NCBI gene identifier: 1288), NO: 9), COL6A2 gene (see, e.g., NCBI gene identifier: 1292), COL6A3 gene (see, e.g., NCBI gene identifier: 1293), COL6A4 gene (see, e.g., NCBI gene identifier: 344875), COL6A5 gene (see, e.g., NCBI gene identifier: 256076), COL6A6 gene (see, e.g., NCBI gene identifier: 131873), COL7A1 gene (see, e.g., NCBI gene identifier: 1294;SEQ ID NO: 10), COL8A1 gene (see, e.g., NCBI gene identifier: 1295), COL9A1 gene (see, e.g., NCBI gene identifier: 1297), COL9A2 gene (see, e.g., NCBI gene identifier: 1298), COL9A3 gene (see, e.g., NCBI gene identifier: 1299), COL10A1 gene (see, e.g., NCBI gene identifier: 1300), COL11A1 gene (see, e.g., NCBI gene identifier: 1301), COL11A2 gene (see, e.g., NCBI gene identifier: 1302), NCBI gene identifier: 1302), COL12A1 gene (see, e.g., NCBI gene identifier: 1303), COL13A1 gene (see, e.g., NCBI gene identifier: 1305), COL14A1 gene (see, e.g., NCBI gene identifier: 7373), COL15A1 gene (see, e.g., NCBI gene identifier: 1306), COL16A1 gene (see, e.g., NCBI gene identifier: 1307), COL17A1 gene (see, e.g., NCBI gene identifier: 1308; SEQ ID NO: 1310). , COL2A1 gene (see, e.g., NCBI Gene Identifier: 81578), COL22A1 gene (see, e.g., NCBI Gene Identifier: 169044), COL23A1 gene (see, e.g., NCBI Gene Identifier: 91522), COL24A1 gene (see, e.g., NCBI Gene Identifier: 255631), COL25A1 gene (see, e.g., NCBI Gene Identifier: 84570), COL26A1 gene (see, e.g., NCBI Gene Identifier: 136227), COL27A1 gene (see, e.g., NCBI Gene Identifier: 85301), COL28A1 gene (see, e.g., NCBI Gene ID: 340267), etc. In some embodiments, the polynucleotides of the present disclosure (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any human collagen gene (and / or its coding sequence) described herein or known in the art.;

[0128] In some embodiments, the polynucleotides of the present disclosure (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprise codon-optimized variants of any human collagen gene described herein. In some embodiments, the use of codon-optimized variants of human collagen genes increases the stability and / or yield of heterologous expression (RNA and / or protein) of human collagen in target cells (such as human keratinocytes or fibroblasts) compared to the stability and / or yield of heterologous expression of corresponding non-codon-optimized wild-type sequences.

[0129] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL1A1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

[0130] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 1 or SEQ ID NO: 2 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000 but less than 4395 consecutive nucleotides of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-4392 of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-4392 of SEQ ID NO: 1 or SEQ ID NO: 2.

[0131] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL1A2 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 3 or SEQ ID NO: 4. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 3 or SEQ ID NO: 4.

[0132] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 3 or SEQ ID NO: 4. In some embodiments, the 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 3 or SEQ ID NO: 4 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000, but less than 4101 consecutive nucleotides of SEQ ID NO: 3 or SEQ ID NO: 4. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-4098 of SEQ ID NO: 3 or SEQ ID NO: 4. In some embodiments, the polynucleotides of the present disclosure comprise a sequence of nucleic acids 1-4098 of SEQ ID NO: 3 or SEQ ID NO: 4.

[0133] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL3A1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 5 or SEQ ID NO: 6.

[0134] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 5 or SEQ ID NO: 6 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000 but less than 4401 consecutive nucleotides of SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-4398 of SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-4398 of SEQ ID NO: 5 or SEQ ID NO: 6.

[0135] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL4A1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 7 or SEQ ID NO: 8. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 7 or SEQ ID NO: 8.

[0136] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 7 or SEQ ID NO: 8. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 7 or SEQ ID NO: 8 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000, at least 4500, at least 5000, but less than 5010 consecutive nucleotides of SEQ ID NO: 7 or SEQ ID NO: 8. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-5007 of SEQ ID NO: 7 or SEQ ID NO: 8. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-5007 of SEQ ID NO: 7 or SEQ ID NO: 8.

[0137] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL6A1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 9 or SEQ ID NO: 10. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 9 or SEQ ID NO: 10.

[0138] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 9 or SEQ ID NO: 10. In some embodiments, the 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 9 or SEQ ID NO: 10 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, but less than 3087 consecutive nucleotides of SEQ ID NO: 9 or SEQ ID NO: 10. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-3084 of SEQ ID NO: 9 or SEQ ID NO: 10. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-3084 of SEQ ID NO: 9 or SEQ ID NO: 10.

[0139] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL7A1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 11 or SEQ ID NO: 12. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 11 or SEQ ID NO: 12.

[0140] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 11 or SEQ ID NO: 12. In some embodiments, the 5' truncated form, 3' truncated form or fragment of the sequence of SEQ ID NO: 11 or SEQ ID NO: 12 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000, at least 4500, at least 5000, at least 5500, at least 6000, at least 6500, at least 7000, at least 7500, at least 8000, at least 8500 but less than 8835 consecutive nucleotides of SEQ ID NO: 11 or SEQ ID NO: 12. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-8832 of SEQ ID NO: 11 or SEQ ID NO: 12. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-8832 of SEQ ID NO: 11 or SEQ ID NO: 12.

[0141] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human COL17A1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 13 or SEQ ID NO: 14. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 13 or SEQ ID NO: 14.

[0142] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 13 or SEQ ID NO: 14. In some embodiments, the 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 13 or SEQ ID NO: 14 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000 but less than 4494 consecutive nucleotides of SEQ ID NO: 13 or SEQ ID NO: 14. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-4491 of SEQ ID NO: 13 or SEQ ID NO: 14. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-4491 of SEQ ID NO: 13 or SEQ ID NO: 14.

[0143] In some embodiments, the polynucleotides encoding one or more human collagens (e.g., a first human collagen, another human collagen, an additional human collagen, and / or a second human collagen) of the present disclosure have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14. In some embodiments, the polynucleotides encoding one or more human collagens (e.g., a first human collagen, another human collagen, an additional human collagen, and / or a second human collagen) of the present disclosure comprise a sequence selected from SEQ ID NOs: 1-14.

[0144] Polynucleotide encoding fibronectin

[0145] In some embodiments, the disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of a fibronectin gene. The coding sequence of any fibronectin gene (including any subtype thereof) from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, human fibronectin gene (see, for example, NCBI gene identifier: 2335), mouse fibronectin gene (see, for example, NCBI gene identifier: 14268), chimpanzee fibronectin gene (see, for example, NCBI gene identifier: 459926), rat fibronectin gene (see, for example, NCBI gene identifier: 25661), rabbit fibronectin gene (see, for example, NCBI gene identifier: 100328589), etc. Methods for identifying fibronectin gene homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of any fibronectin gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure comprise a codon optimized variant of any fibronectin gene (and / or its coding sequence) described herein or known in the art.

[0146] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising the coding sequence of a human fibronectin gene. In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of a human FN1 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 35 or SEQ ID NO: 36. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 35 or SEQ ID NO: 36.

[0147] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO:35 or SEQ ID NO:36. In some embodiments, a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO:35 or SEQ ID NO:36 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000, at least about 4500, at least about 5000, at least about 5500, at least about 6000, at least about 6500, at least about 7000, but less than 7434 consecutive nucleotides of SEQ ID NO:35 or SEQ ID NO:36. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-7431 of SEQ ID NO: 35 or SEQ ID NO: 36. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-7431 of SEQ ID NO: 35 or SEQ ID NO: 36.

[0148] Polynucleotide encoding elastin

[0149] In some embodiments, the disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of an elastin gene. The coding sequence of any elastin gene (including any subtype thereof) from any suitable species known in the art, including, for example, human elastin gene (see, for example, NCBI gene identifier: 2006), mouse elastin gene (see, for example, NCBI gene identifier: 13717), chimpanzee elastin gene (see, for example, NCBI gene identifier: 463943), rat elastin gene (see, for example, NCBI gene identifier: 25043), rabbit elastin gene (see, for example, NCBI gene identifier: 100344271), etc., can be encoded by the polynucleotides of the disclosure. Methods for identifying homologs / orthologs of elastin genes from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure comprise sequences having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequences of any elastin gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure comprise codon-optimized variants of any elastin gene (and / or its coding sequence) described herein or known in the art.

[0150] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising the coding sequence of a human elastin gene. In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of a human ELN gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 37 or SEQ ID NO: 38. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 37 or SEQ ID NO: 38.

[0151] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 37 or SEQ ID NO: 38. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 37 or SEQ ID NO: 38 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, but less than 2361 consecutive nucleotides of SEQ ID NO: 37 or SEQ ID NO: 38. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-2358 of SEQ ID NO: 37 or SEQ ID NO: 38. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-2358 of SEQ ID NO: 37 or SEQ ID NO: 38.

[0152] Polynucleotide encoding a photoreceptin protein

[0153] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of a lumican gene. The coding sequence of any lumican gene (including any subtype thereof) from any suitable species known in the art, including, for example, human lumican gene (see, for example, NCBI gene identifier: 4060), mouse lumican gene (see, for example, NCBI gene identifier: 17022), chimpanzee lumican gene (see, for example, NCBI gene identifier: 452119), rat lumican gene (see, for example, NCBI gene identifier: 81682), rabbit lumican gene (see, for example, NCBI gene identifier: 100008665), etc., can be encoded by the polynucleotides of the present disclosure. Methods for identifying lumican gene homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of any aphrodisiac gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure comprise a codon-optimized variant of any aphrodisiac gene (and / or its coding sequence) described herein or known in the art.

[0154] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising the coding sequence of a human aureoglycan gene. In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of a human LUM gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 39 or SEQ ID NO: 40. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 39 or SEQ ID NO: 40.

[0155] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 39 or SEQ ID NO: 40. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 39 or SEQ ID NO: 40 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, but less than 1017 consecutive nucleotides of SEQ ID NO: 39 or SEQ ID NO: 40. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-1014 of SEQ ID NO: 39 or SEQ ID NO: 40. In some embodiments, the polynucleotides of the present disclosure comprise a sequence of nucleic acids 1-1014 of SEQ ID NO: 39 or SEQ ID NO: 40.

[0156] Polynucleotides encoding vitronectin and vitronectin receptor proteins

[0157] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of a vitronectin or vitronectin receptor gene. The coding sequence of any vitronectin or vitronectin receptor gene (including any subtype thereof) from any suitable species known in the art, including, for example, human vitronectin or vitronectin receptor genes (see, for example, NCBI gene identifiers: 7448 and 3685), mouse vitronectin or vitronectin receptor genes (see, for example, NCBI gene identifiers: 22370 and 16410), chimpanzee vitronectin or vitronectin receptor genes (see, for example, NCBI gene identifiers: 738261 and 459807), rat vitronectin or vitronectin receptor genes (see, for example, NCBI gene identifiers: 29169 and 257645), rabbit vitronectin or vitronectin receptor genes (see, for example, NCBI gene identifiers: 100009128 and 100008956), etc. can be encoded by the polynucleotides of the present disclosure. Methods for identifying vitronectin or vitronectin receptor gene homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure comprise sequences having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of any vitronectin or vitronectin receptor gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure comprise codon-optimized variants of any vitronectin or vitronectin receptor gene (and / or its coding sequence) described herein or known in the art.

[0158] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising the coding sequence of human vitronectin or vitronectin receptor genes. In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of human VTN gene (or its codon optimized variant). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 41 or SEQ ID NO: 42. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 41 or SEQ ID NO: 42.

[0159] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 41 or SEQ ID NO: 42. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 41 or SEQ ID NO: 42 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least about 1250 but less than 1437 consecutive nucleotides of SEQ ID NO: 41 or SEQ ID NO: 42. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-1034 of SEQ ID NO: 41 or SEQ ID NO: 42. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-1034 of SEQ ID NO: 41 or SEQ ID NO: 42.

[0160] Polynucleotide encoding laminin

[0161] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of a laminin gene. The coding sequence of any laminin gene (including any isoform thereof) from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, and the laminin gene includes, for example, human laminin gene (see, for example, NCBI gene identifiers: 284217, 3908, 3909, 3910, 3911, 3912, 3913, 3914, 3915, 3918 and 10319), mouse ... BI gene identifiers: 16774, 16780, and 16782), chimpanzee laminin genes (see, e.g., NCBI gene identifiers: 455339, 469668, and 457571), rat laminin genes (see, e.g., NCBI gene identifiers: 307582, 305078, and 192362), rabbit laminin genes (see, e.g., NCBI gene identifiers: 100346886 and 100342905), etc. Methods for identifying laminin gene homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of any laminin gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure comprise a codon-optimized variant of any laminin gene (and / or its coding sequence) described herein or known in the art.

[0162] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising a coding sequence of a human laminin gene, such as a human LAMA1 gene (see, e.g., NCBI gene identifier: 284217), a human LAMA2 gene (see, e.g., NCBI gene identifier: 3908), a human LAMA3 gene (see, e.g., NCBI gene identifier: 3909), a human LAMA4 gene (see, e.g., NCBI gene identifier: 3910), a human LAMA5 gene (see, e.g., NCBI gene identifier: 3911), a human LAMB1 gene (see, e.g., NCBI gene identifier: 3912), a human LAMB2 gene (see, e.g., NCBI gene identifier: 3913), a human LAMB3 gene (see, e.g., NCBI gene identifier: 3914), a human LAMC1 gene (see, e.g., NCBI gene identifier: 3915), a human LAMC2 gene (see, e.g., NCBI gene identifier: 3918), or a human LAMC3 gene (see, e.g., NCBI gene identifier: 10319).

[0163] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human LAMA3 gene (or its codon optimized variant). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 43 or SEQ ID NO: 44. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 43 or SEQ ID NO: 44.

[0164] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 43 or SEQ ID NO: 44. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 43 or SEQ ID NO: 44 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000, at least about 4500, at least about 5000, but less than 5175 consecutive nucleotides of SEQ ID NO: 43 or SEQ ID NO: 44. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-5172 of SEQ ID NO: 43 or SEQ ID NO: 44. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-5172 of SEQ ID NO: 43 or SEQ ID NO: 44.

[0165] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human LAMB3 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 45 or SEQ ID NO: 46. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 45 or SEQ ID NO: 46.

[0166] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 45 or SEQ ID NO: 46. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 45 or SEQ ID NO: 46 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, but less than 3519 consecutive nucleotides of SEQ ID NO: 45 or SEQ ID NO: 46. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-3516 of SEQ ID NO: 45 or SEQ ID NO: 46. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-3516 of SEQ ID NO: 45 or SEQ ID NO: 46.

[0167] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the human LAMC2 gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 47 or SEQ ID NO: 48.

[0168] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 47 or SEQ ID NO: 48 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, but less than 3582 consecutive nucleotides of SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-3579 of SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of nucleic acids 1-3579 of SEQ ID NO: 47 or SEQ ID NO: 48.

[0169] Polynucleotide encoding neuromodulator protein

[0170] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising coding sequences of neuromodulator genes. The coding sequences of any neuromodulator genes (including any subtypes thereof) from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, clostridium botulinum neuromodulator genes (see, for example, NCBI gene identifiers: 5185061 and 39483740), etc. Methods for identifying homologs / orthologs of neuromodulator genes from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of any neuromodulator gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure comprise a codon-optimized variant of any neuromodulator gene (and / or its coding sequence) described herein or known in the art.

[0171] In some embodiments, the disclosure relates to one or more polynucleotides (ie, one or more first polynucleotides and / or one or more second polynucleotides) comprising a coding sequence for a Clostridium botulinum neuromodulator gene.

[0172] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the Clostridium botulinum botA gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 49 or SEQ ID NO: 50. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 49 or SEQ ID NO: 50.

[0173] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 49 or SEQ ID NO: 50. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 49 or SEQ ID NO: 50 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least about 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, but less than 3891 consecutive nucleotides of SEQ ID NO: 49 or SEQ ID NO: 50. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-3888 of SEQ ID NO: 49 or SEQ ID NO: 50. In some embodiments, the polynucleotides of the present disclosure comprise a sequence of nucleic acids 1-3888 of SEQ ID NO: 49 or SEQ ID NO: 50.

[0174] In some embodiments, the polynucleotides of the present disclosure comprise the coding sequence of the Clostridium botulinum botB gene (or a codon-optimized variant thereof). In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 51 or SEQ ID NO: 52. In some embodiments, the polynucleotides of the present disclosure comprise the sequence of SEQ ID NO: 51 or SEQ ID NO: 52.

[0175] In some embodiments, the polynucleotides of the present disclosure comprise a 5' truncated form, a 3' truncated form, or a fragment of the sequence of SEQ ID NO: 51 or SEQ ID NO: 52. In some embodiments, the 5' truncated form, the 3' truncated form, or a fragment of the sequence of SEQ ID NO: 51 or SEQ ID NO: 52 is a polynucleotide having at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least about 1250, at least 1500, at least 1750, at least 2000, at least 2500, at least 3000, at least 3500, but less than 3876 consecutive nucleotides of SEQ ID NO: 51 or SEQ ID NO: 52. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of nucleic acids 1-3873 of SEQ ID NO: 51 or SEQ ID NO: 52. In some embodiments, the polynucleotides of the present disclosure comprise a sequence of nucleic acids 1-3873 of SEQ ID NO: 51 or SEQ ID NO: 52.

[0176] Polynucleotide encoding fibrillin

[0177] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising one or more polynucleotides comprising a coding sequence of a fibrillin gene. The coding sequence of any fibrillin gene (including any isoform thereof) from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, human fibrillin gene (see, for example, NCBI gene identifiers: 2200, 2201, and 84467), mouse fibrillin gene (see, for example, NCBI gene identifiers: 14118 and 14119), chimpanzee fibrillin gene (see, for example, NCBI gene identifiers: 453411, 471621, and 455669), rat fibrillin gene (see, for example, NCBI gene identifiers: 83727 and 689008), rabbit fibrillin gene (see, for example, NCBI gene identifiers: 100350931, 100357126, and 100359336), etc. Methods for identifying the fibrillin gene homologues / orthologues from additional species are known to those of ordinary skill in the art. In some embodiments, the polynucleotides of the present disclosure include sequences with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to any fibrillin gene (and / or its coding sequence) described herein or known in the art. In some embodiments, the polynucleotides of the present disclosure include codon optimized variants of any fibrillin gene (and / or its coding sequence) described herein or known in the art.

[0178] In some embodiments, the present disclosure relates to one or more polynucleotides (i.e., one or more first polynucleotides and / or one or more second polynucleotides) comprising a coding sequence of a human fibrillin gene, such as the human FBN1 gene (see, e.g., NCBI gene identifier: 2200), the human FBN2 gene (see, e.g., NCBI gene identifier: 2201), or the human FBN3 gene (see, e.g., NCBI gene identifier: 84467).

[0179] Exemplary polynucleotides

[0180] In some embodiments, the polynucleotides encoding one or more cosmetic proteins (e.g., the first cosmetic protein, another cosmetic protein, an additional cosmetic protein, and / or a second cosmetic protein) of the present disclosure have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14 or 35-52. In some embodiments, the polynucleotides encoding one or more cosmetic proteins (e.g., the first cosmetic protein, another cosmetic protein, an additional cosmetic protein, and / or a second cosmetic protein) of the present disclosure comprise a sequence selected from SEQ ID NOs: 1-14 or 35-52.

[0181] In some embodiments, the polynucleotides encoding one or more cosmetic proteins (e.g., the first cosmetic protein, another cosmetic protein, an additional cosmetic protein, and / or a second cosmetic protein) of the present disclosure have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOs: 1-14, 35-38, or 43-48. In some embodiments, the polynucleotides encoding one or more cosmetic proteins (e.g., the first cosmetic protein, another cosmetic protein, an additional cosmetic protein, and / or a second cosmetic protein) of the present disclosure comprise a sequence selected from SEQ ID NOs: 1-14, 35-38, or 43-48.

[0182] The polynucleotides encoding beauty proteins (e.g., human collagen) disclosed in the present invention may also encode additional coding sequences and non-coding sequences. Examples of additional coding sequences and non-coding sequences may include, but are not limited to, sequences encoding additional polypeptide tags (e.g., in-frame coding with beauty proteins to produce fusion proteins), introns (e.g., natural, modified or heterologous introns), 5' and / or 3'UTRs (e.g., natural, modified or heterologous 5' and / or 3'UTRs), etc. Suitable examples of polypeptide tags may include, but are not limited to, purification tags, such as his tags, flag tags, maltose binding protein, and glutathione-S-transferase tags; detection tags, such as tags that can be detected photometrically (e.g., green fluorescent protein, red fluorescent protein, etc.); and tags with detectable enzymatic activity (e.g., alkaline phosphatase, etc.); tags containing secretion sequences, signal sequences, leader sequences, and / or stabilization sequences; any combination of protease cleavage sites (e.g., furin cleavage sites, TEV cleavage sites, thrombin cleavage sites, etc.), etc. In some embodiments, the 5' and / or 3'UTR increases the stability, localization and / or translation efficiency of the polynucleotide. In some embodiments, the 5' and / or 3'UTR improves the level and / or duration of protein expression. In some embodiments, the 5' and / or 3'UTR includes elements (e.g., one or more miRNA binding sites, etc.) that can block or reduce off-target expression (e.g., inhibit expression in specific cell types (e.g., neuronal cells), at specific times in the cell cycle, at specific developmental stages, etc.). In some embodiments, the 5' and / or 3'UTR includes elements (e.g., one or more miRNA binding sites, etc.) that can enhance the expression of beauty proteins in specific cell types (such as human keratinocytes and / or fibroblasts).

[0183] In some embodiments, the polynucleotide encoding the beauty protein (e.g., human collagen) of the present disclosure is operably linked to one or more (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.) regulatory sequences. The term "regulatory sequence" may include enhancers, insulators, promoters, and other expression control elements (e.g., polyadenylation signals). Any one or more suitable enhancers known in the art may be used, including, for example, enhancer sequences from mammalian genes (such as globulin, elastase, albumin, α-fetoprotein, insulin, etc.), enhancer sequences from eukaryotic cell viruses (such as the SV40 enhancer (bp100-270) on the rear side of the replication origin, cytomegalovirus early promoter enhancer, polyoma enhancer on the rear side of the replication origin, adenovirus enhancer, etc.), and any combination thereof. Any one or more suitable insulators known in the art may be used, including, for example, the HSV chromatin boundary (CTRL / CTCF binding / insulator) elements CTRL1 and / or CTRL2, the chicken hypersensitive site 4 insulator (cHS4), the human HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE), the scaffold / matrix attachment region (S / MAR) from the human interferon beta gene (IFNB1), and any combination thereof. Any suitable promoter known in the art (e.g., suitable for transcription in a mammalian host cell) may be used, including, for example, promoters obtained from the genome of a virus (such as polyoma virus, fowlpox virus, adenovirus (such as adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, retrovirus, hepatitis B virus, Simian virus 40 (SV40), etc.), promoters from heterologous mammalian genes (such as actin promoters (e.g., β-actin promoter), ubiquitin promoters (e.g., ubiquitin C (UbC) promoter), phosphoglycerate kinase (PGK) promoter, immunoglobulin promoters, from heat shock promoters, etc.), promoters from homologous mammalian genes (e.g., native human collagen, fibronectin, elastin, alginate, vitronectin, laminin and / or fibrillin promoters), synthetic promoters (such as the CAGG promoter), and any combination thereof, provided that the promoter is compatible with the host cell. Regulatory sequences may include those that direct constitutive expression of the nucleic acid, as well as tissue-specific regulatory sequences and / or inducible or repressible sequences.

[0184] In some embodiments, the polynucleotides encoding beauty proteins (e.g., human collagen) of the present disclosure are operably linked to one or more heterologous promoters. In some embodiments, one or more heterologous promoters are constitutive promoters, tissue-specific promoters, temporal promoters, spatial promoters, inducible promoters, and repressible promoters. One or more. In some embodiments, one or more heterologous promoters are human cytomegalovirus (HCMV) immediate early promoter, human elongation factor-1 (EF1) promoter, human β-actin promoter, human UbC promoter, human PGF promoter, synthetic CAGG promoter, and any combination thereof. In some embodiments, the polynucleotides encoding beauty proteins (e.g., human collagen) of the present disclosure are operably linked to HCMV promoters.

[0185] In some embodiments, the polynucleotides of the present disclosure do not comprise a coding sequence for a collagen alpha-1 (VII) chain polypeptide (COL7), such as a transgene encoding a collagen alpha-1 (VII) chain polypeptide (COL7). In some embodiments, the polynucleotides of the present disclosure do not comprise a coding sequence for a lysyl hydroxylase 3 polypeptide (LH3), such as a transgene encoding a lysyl hydroxylase 3 polypeptide (LH3). In some embodiments, the polynucleotides of the present disclosure do not comprise a coding sequence for a keratin type I cytoskeleton 17 polypeptide (KRT17), such as a transgene encoding a keratin type I cytoskeleton 17 polypeptide (KRT17). In some embodiments, the polynucleotides of the present disclosure do not comprise a coding sequence for a transglutaminase (TGM) polypeptide (e.g., a human transglutaminase polypeptide, such as a human TGM1 polypeptide), such as a transgene encoding a transglutaminase (TGM) polypeptide (e.g., a human transglutaminase polypeptide, such as a human TGM1 polypeptide). In some embodiments, the polynucleotides of the present disclosure do not comprise a coding sequence for a laminin subunit beta-3 polypeptide (LAMB3), such as a transgene encoding a laminin subunit beta-3 polypeptide (LAMB3). In some embodiments, the polynucleotides of the present disclosure do not comprise a coding sequence for a collagen αt-1 (VII) chain polypeptide, a lysyl hydroxylase 3 polypeptide, a keratin type I cytoskeleton 17 polypeptide, and / or any chimeric polypeptides thereof, such as a transgene encoding a collagen αt-1 (VII) chain polypeptide, a lysyl hydroxylase 3 polypeptide, a keratin type I cytoskeleton 17 polypeptide, and / or any chimeric polypeptides thereof. In some embodiments, the polynucleotides of the present disclosure do not comprise coding sequences for collagen alpha-1 (VII) chain polypeptides, lysyl hydroxylase 3 polypeptides, keratin type I cytoskeleton 17 polypeptides, transglutaminase (TGM) polypeptides (e.g., human transglutaminase polypeptides, such as human TGM1 polypeptides), laminin subunit beta-3 (LAMB3) polypeptides (e.g., human LamB3 polypeptides), and / or any chimeric polypeptides thereof (e.g., transgenes encoding collagen alpha-1 (VII) chain polypeptides, lysyl hydroxylase 3 polypeptides, keratin type I cytoskeleton 17 polypeptides, transglutaminase (TGM) polypeptides (e.g., human transglutaminase polypeptides, such as human TGM1 polypeptides), laminin subunit alpha-3 (LAMB3) polypeptides (e.g., human LamB3 polypeptides), and / or any chimeric polypeptides thereof).

[0186] Beauty Protein

[0187] Collagen

[0188] In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length collagen or any subtype or part thereof. Any collagen from any suitable species known in the art can be encoded by the polynucleotides disclosed herein, including, for example, human collagen (see, for example, UniProt accession numbers P02452, P08123, P02461, P02462, P08572, P12109, Q02388, Q9UMD9, etc.), mouse collagen (see, for example, UniProt accession numbers P11087, Q01149, P08121, P02463, P08122, Q04857, Q63870, Q07563, etc.), chimpanzee collagen (see, for example, UniProt accession numbers P11087, Q01149, P08121, P02463, P08122, Q04857, Q63870, Q07563, etc.), 4, etc.), rat collagen (see, e.g., UniProt Accession Nos. A0A2I3SM98, A0A2J8L483, H2QJ46, K7C8P4, K7C8W0, A0A2J8M8U9, H2QMJ5, H2Q2J4, etc.), rat collagen (see, e.g., UniProt Accession Nos. P02454, P02466, P13941, P02466, F1M6Q3, D3ZUL3, D3ZE04, D3ZE04, etc.), rabbit collagen (see, e.g., UniProt Accession Nos. G1T4A5, Q28668, G1T8J0, G1U9R7, G1T548, G1T380, G1T548, etc.), etc. Methods for identifying collagen homologs / orthologs from additional species are known to those of ordinary skill in the art, including, for example, using amino acid sequence alignment programs such as blastp suite or OrthoDB. In some embodiments, the collagen polypeptides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any collagen polypeptide described herein or known in the art.

[0189] In some embodiments, the present disclosure relates to one or more polynucleotides encoding human collagen. Any suitable human collagen known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, collagen alpha-1 (I) chain polypeptide (COL1-1) (see, e.g., UniProt Accession No. P02452; SEQ ID NO: 15), collagen alpha-2 (I) chain polypeptide (COL1-2) (see, e.g., UniProt Accession No. P08123; SEQ ID NO: 16), collagen alpha-1 (II) chain polypeptide (COL2) (see, e.g., UniProt Accession No. P02458), collagen alpha-1 (III) chain polypeptide (COL3) (see, e.g., UniProt Accession No. P2461; SEQ ID NO: 17), collagen alpha-1 (IV) chain polypeptide (COL4-1) (see, e.g., UniProt Accession No. P02462; SEQ ID NO: 18), collagen alpha-1 (III) chain polypeptide (COL4-2) (see, e.g., UniProt Accession No. P02463; SEQ ID NO: 19), collagen alpha-1 (III) chain polypeptide (COL5-3) (see, e.g., UniProt Accession No. P02464; SEQ ID NO: 20), collagen alpha-1 (III) chain polypeptide (COL6-4) (see, e.g., UniProt Accession No. P02465; SEQ ID NO: 21), collagen alpha-1 (III) chain polypeptide (COL7-5) (see, e.g., UniProt Accession No. P02467; SEQ ID NO: 22), collagen alpha-1 (III) chain polypeptide (COL8-6) (see, e.g., UniProt Accession No. P02468; SEQ ID NO: 23), NO: 18), collagen alpha-2 (IV) chain polypeptide (COL4-2) (see, e.g., UniProt Accession No. P08572), collagen alpha-3 (IV) chain polypeptide (COL4-3) (see, e.g., UniProt Accession No. Q01955), collagen alpha-4 (IV) chain polypeptide (COL4-4) (see, e.g., UniProt Accession No. P53420), collagen alpha-5 (IV) chain polypeptide (COL4-5) (see, e.g., UniProt Accession No. 29400), collagen alpha-6 (IV) chain polypeptide (COL4-6) (see, e.g., UniProt Accession No. 29401), collagen alpha-7 (IV) chain polypeptide (COL4-7) (see, e.g., UniProt Accession No. 29403), collagen alpha-8 (IV) chain polypeptide (COL4-8) (see, e.g., UniProt Accession No. 29404), collagen alpha-9 (IV) chain polypeptide (COL4-10) (see, e.g., UniProt Accession No. 29405), collagen alpha-10 (IV) chain polypeptide (COL4-11) (see, e.g., UniProt Accession No. 29406), collagen alpha-11 (IV) chain polypeptide (COL4-12) (see, e.g., UniProt Accession No. 29407), collagen alpha-12 (IV) chain polypeptide (COL4-13) (see, e.g., UniProt Accession No. 29408), collagen alpha-13 (IV) chain polypeptide (COL4-14) (see, e.g., UniProt Accession No. 29409), collagen alpha-14 (IV) chain polypeptide (COL4-15) (see, e.g., Uni 6) (see, e.g., UniProt Accession No. Q14031), collagen alpha-1 (V) chain polypeptide (COL5-1) (see, e.g., UniProt Accession No. P20908), collagen alpha-2 (V) chain polypeptide (COL5-2) (see, e.g., UniProt Accession No. P05997), collagen alpha-3 (V) chain polypeptide (COL5-3) (see, e.g., UniProt Accession No. P25940), collagen alpha-1 (VI) chain polypeptide (COL6-1) (see, e.g., UniProt Accession No. P12109; SEQ ID NO: 514); ID NO: 19), collagen alpha-2 (VI) chain polypeptide (COL6-2) (see, e.g., UniProt Accession No. P12110), collagen alpha-3 (VI) chain polypeptide (COL6-3) (see, e.g., UniProt Accession No. P12111), collagen alpha-4 (VI) chain polypeptide (COL6-4), collagen alpha-5 (VI) chain polypeptide (COL6-5) (see, e.g., UniProt Accession No. A8TX70), collagen alpha-6 (VI) chain polypeptide (COL6-6) (see, e.g., UniProt Accession No. A6NMZ7), collagen alpha-1 (VII) chain polypeptide (COL7) (see, e.g., UniProt Accession No. Q02388;SEQ ID NO: 20), collagen alpha-1 (VIII) chain polypeptide (COL8) (see, e.g., UniProt Accession No. P27658), collagen alpha t-1 (IX) chain polypeptide (COL9-1) (see, e.g., UniProt Accession No. P20849), collagen alpha-2 (IX) chain polypeptide (COL9-2) (see, e.g., UniProt Accession No. Q14055), collagen alpha-3 (IX) chain polypeptide (COL9-3) (see, e.g., UniProt Accession No. Q14050), collagen alpha-1 (X) chain polypeptide (COL10) (see, e.g., UniProt Accession No. Q03692), collagen alpha-1 (XI) chain polypeptide (COL11-1) (see, e.g., UniProt Accession No. P12107), collagen alpha-2 (XI) chain polypeptide (COL11-2) (see, e.g., UniProt Accession No. Q14057), collagen alpha-3 (IX) chain polypeptide (COL11-3) (see, e.g., UniProt Accession No. Q14058), collagen alpha-1 (X) chain polypeptide (COL10) (see, e.g., UniProt Accession No. Q03692), collagen alpha-1 (XI) chain polypeptide (COL11-1) (see, e.g., UniProt Accession No. P12107), collagen alpha-2 (XI) chain polypeptide (COL11 -2) (see, e.g., UniProt Accession No. P13942), collagen alpha-1 (XII) chain polypeptide (COL12) (see, e.g., UniProt Accession No. Q99715), collagen alpha-1 (XIII) chain polypeptide (COL13) (see, e.g., UniProt Accession No. Q5TAT6), collagen alpha-1 (XIV) chain polypeptide (COL14) (see, e.g., UniProt Accession No. Q05707), collagen alpha-1 (XV) chain polypeptide (COL15) (see, e.g., UniProt Accession No. P39059), collagen alpha-1 (XVI) chain polypeptide (COL16) (see, e.g., UniProt Accession No. Q07092), collagen alpha-1 (XVII) chain polypeptide (COL17) (see, e.g., UniProt Accession No. Q9UMD9;SEQ ID NO: 21), collagen alpha-1 (XVIII) chain polypeptide (COL18) (see, e.g., UniProt Accession No. P39060), collagen alpha-1 (XIX) chain polypeptide (COL19) (see, e.g., UniProt Accession No. Q14993), collagen alpha-1 (XX) chain polypeptide (COL20) (see, e.g., UniProt Accession No. Q9P218), collagen alpha-1 (XXI) chain polypeptide (COL21) (see, e.g., UniProt Accession No. Q96P44), collagen alpha-1 (XXII) chain polypeptide (COL22) (see, e.g., UniProt Accession No. Q8NFW1), collagen alpha-1 (XXIII) chain polypeptide (COL23) (See, e.g., UniProt Accession No. Q86Y22), collagen alpha-1 (XXIV) chain polypeptide (COL24) (See, e.g., UniProt Accession No. Q17RW2), collagen alpha-1 (XXV) chain polypeptide (COL25) (See, e.g., UniProt Accession No. Q9BXS0), collagen alpha-1 (XXVI) chain polypeptide (COL26) (See, e.g., UniProt Accession No. Q96A83), collagen alpha-1 (XXVII) chain polypeptide (COL27) (See, e.g., UniProt Accession No. Q8IZC6), collagen alpha-1 (XXVIII) chain polypeptide (COL28) (See, e.g., UniProt Accession No. Q2UY09), etc. In some embodiments, the polynucleotides of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence encoding any human collagen polypeptide described herein or known in the art. Methods for identifying additional human collagen or collagen-like polypeptide homologs / orthologs are known to those of ordinary skill in the art and include, for example, the use of amino acid sequence alignment programs such as; blastp suite or OrthoDB.

[0190] In some embodiments, the polynucleotides of the present disclosure encode human COL1-1 protein. In some embodiments, the polynucleotides encoding COL1-1 protein are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 15. In some embodiments, the polynucleotides encoding human COL1-1 protein are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 15.

[0191] In some embodiments, the polynucleotide encoding the COL1-1 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 15. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400 but less than 1464 consecutive amino acids of SEQ ID NO: 15.

[0192] In some embodiments, the polynucleotides of the present disclosure encode human COL1-2 proteins. In some embodiments, the polynucleotides encoding COL1-2 proteins are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 16. In some embodiments, the polynucleotides encoding human COL1-2 proteins are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 16.

[0193] In some embodiments, the polynucleotide encoding the COL1-2 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 16. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300 but less than 1366 consecutive amino acids of SEQ ID NO: 16.

[0194] In some embodiments, the polynucleotides of the present disclosure encode human COL3 proteins. In some embodiments, the polynucleotides encoding COL3 proteins are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 17. In some embodiments, the polynucleotides encoding human COL3 proteins are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 17.

[0195] In some embodiments, the polynucleotide encoding the COL3 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 17. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400 but less than 1466 consecutive amino acids of SEQ ID NO: 17.

[0196] In some embodiments, the polynucleotides of the present disclosure encode human COL4-1 protein. In some embodiments, the polynucleotide encoding the COL4-1 protein is a polynucleotide encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 18. In some embodiments, the polynucleotide encoding the human COL4-1 protein is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 18.

[0197] In some embodiments, the polynucleotide encoding the COL4-1 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 18. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600 but less than 1669 consecutive amino acids of SEQ ID NO: 18.

[0198] In some embodiments, the polynucleotides of the present disclosure encode human COL6-1 protein. In some embodiments, the polynucleotides encoding COL6-1 protein are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 19. In some embodiments, the polynucleotides encoding human COL6-1 protein are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 19.

[0199] In some embodiments, the polynucleotide encoding the COL6-1 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 19. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000 but less than 1028 consecutive amino acids of SEQ ID NO: 19.

[0200] In some embodiments, the polynucleotides of the present disclosure encode human COL7 proteins. In some embodiments, the polynucleotides encoding COL7 proteins are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 20. In some embodiments, the polynucleotides encoding human COL7 proteins are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 20.

[0201] In some embodiments, the polynucleotide encoding the COL7 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO:20. 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, at least 2500, at least 2600, at least 2700, at least 2800, at least 2900 but less than 2944 consecutive amino acids of SEQ ID NO:20.

[0202] In some embodiments, the polynucleotides of the present disclosure encode human COL17 proteins. In some embodiments, the polynucleotides encoding COL17 proteins are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 21. In some embodiments, the polynucleotides encoding human COL17 proteins are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 21.

[0203] In some embodiments, the polynucleotide encoding the COL17 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 21. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400 but less than 1497 consecutive amino acids of SEQ ID NO: 21.

[0204] In some embodiments, one or more human collagens of the present disclosure (e.g., a first human collagen, another human collagen, an additional human collagen, and / or a second human collagen) comprise an amino acid sequence comprising at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 15-21. In some embodiments, one or more human collagens of the present disclosure (e.g., a first human collagen, another human collagen, an additional human collagen, and / or a second human collagen) comprise a sequence selected from SEQ ID NOs: 15-21.

[0205] In some embodiments, one or more human collagens of the present disclosure (e.g., a first human collagen, another human collagen, an additional human collagen, and / or a second human collagen) comprise an amino acid sequence comprising at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 15-17. In some embodiments, one or more human collagens of the present disclosure (e.g., a first human collagen, another human collagen, an additional human collagen, and / or a second human collagen) comprise a sequence selected from SEQ ID NOs: 15-17.

[0206] Fibronectin

[0207] In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length fibronectin or any subtype or part thereof. Any fibronectin from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, human fibronectin (see, for example, UniProt accession number P02751), mouse fibronectin (see, for example, UniProt accession number P11276), chimpanzee fibronectin (see, for example, UniProt accession number P11276), rat fibronectin (see, for example, UniProt accession number P04937), rabbit fibronectin (see, for example, UniProt accession number P04937), etc. The method of identifying fibronectin homologues / orthologues from additional species is known to those of ordinary skill in the art. In some embodiments, the fibronectin of the present disclosure comprises a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any fibronectin described herein or known in the art.

[0208] In some embodiments, the polynucleotides of the present disclosure encode human fibronectin. In some embodiments, the polynucleotides encoding human fibronectin are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the sequence of SEQ ID NO: 53. In some embodiments, the polynucleotides encoding human fibronectin are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 53.

[0209] In some embodiments, the polynucleotide encoding human fibronectin is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 53. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400 but less than 2477 consecutive amino acids of SEQ ID NO: 53.

[0210] Elastin and related proteins

[0211] Elastic fibers in the extracellular matrix give elastic properties to tissues. Elastic fibers usually contain two morphologically different components - mature elastin fibers and microfibrils, which mainly contain fibrillin and are associated with other proteins, such as microfibril-associated glycoprotein (MAGP), fibrin and elastin-microfibril interface localization protein (EMILIN). Elastin and its soluble precursor tropoelastin are major structural proteins of the body.

[0212] In some embodiments, the disclosure relates to one or more polynucleotides encoding elastin or elastin-related proteins, and the elastin-related proteins include tropoelastin, fibrillin, microfibril-related glycoprotein, fibrin or elastin-microfibril interface localization protein. In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length elastin or any subtype or part thereof. Any elastin from any suitable species known in the art can be encoded by the polynucleotides of the disclosure, and the elastin includes, for example, human elastin (see, for example, UniProt accession number P15502), mouse elastin (see, for example, UniProt accession number P15502), chimpanzee elastin (see, for example, UniProt accession number H2QUQ6), rat elastin (see, for example, UniProt accession number Q99372) etc. The method for identifying the elastin homologues / orthologues from additional species is known to those of ordinary skill in the art. In some embodiments, the elastin proteins of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any elastin protein described herein or known in the art.

[0213] In some embodiments, the polynucleotides of the present disclosure encode human elastin. In some embodiments, the polynucleotides encoding human elastin are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 54. In some embodiments, the polynucleotides encoding human elastin are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 54.

[0214] In some embodiments, the polynucleotide encoding human elastin is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 54. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700 but less than 786 consecutive amino acids of SEQ ID NO: 54.

[0215] Luminogen activator protein

[0216] In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length alucoprotein or any isoform or portion thereof. Any alucoprotein from any suitable species known in the art may be encoded by the polynucleotides of the disclosure, including, for example, human alucoprotein (see, for example, UniProt Accession No. P51884), mouse alucoprotein (see, for example, UniProt Accession No. P51885), chimpanzee alucoprotein (see, for example, UniProt Accession No. H2Q6L3), rat alucoprotein (see, for example, UniProt Accession No. H2Q6L3), rabbit alucoprotein (see, for example, UniProt Accession No. O46379), etc. Methods for identifying alucoprotein homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the aureoglycan protein of the present disclosure comprises a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any aureoglycan protein described herein or known in the art.

[0217] In some embodiments, the polynucleotides of the present disclosure encode human aureoglycan protein. In some embodiments, the polynucleotide encoding human aureoglycan protein is a polynucleotide encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 55. In some embodiments, the polynucleotide encoding human aureoglycan protein is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 55.

[0218] In some embodiments, the polynucleotide encoding human aureoglycan protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 55. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300 but less than 338 consecutive amino acids of SEQ ID NO: 55.

[0219] Vitronectin and Vitronectin Receptor Protein

[0220] In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length vitronectin or vitronectin receptor proteins or any subtypes or portions thereof. Any vitronectin or vitronectin receptor proteins from any suitable species known in the art may be encoded by the polynucleotides of the disclosure, including, for example, human vitronectin or vitronectin receptor proteins (see, for example, UniProt accession numbers P04004 and P06756), mouse vitronectin or vitronectin receptor proteins (see, for example, UniProt accession numbers P29788 and P43406), chimpanzee vitronectin or vitronectin receptor proteins (see, for example, UniProt accession numbers H2QCH3 and H2R6C3), rat vitronectin or vitronectin receptor proteins (see, for example, UniProt accession number Q7TQ11), rabbit vitronectin or vitronectin receptor proteins (see, for example, UniProt accession number P22458), etc. Methods for identifying vitronectin or vitronectin receptor protein homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the vitronectin or vitronectin receptor protein of the present disclosure comprises a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of any vitronectin or vitronectin receptor protein described herein or known in the art.

[0221] In some embodiments, the polynucleotides of the present disclosure encode human vitronectin. In some embodiments, the polynucleotide encoding human vitronectin is a polynucleotide encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 56. In some embodiments, the polynucleotide encoding human vitronectin is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 56.

[0222] In some embodiments, the polynucleotide encoding human vitronectin is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 56. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, but less than 478 consecutive amino acids of SEQ ID NO: 56.

[0223] Laminin

[0224] In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length laminins or any hypotype thereof or part.Any laminin from any suitable species as known in the art can be encoded by polynucleotides of the present disclosure, and the laminin includes for example human laminin (see for example UniProt accession number P25391, P24043, Q16787, Q16363, O15230, P07942, P55268, Q13751, P11047, Q13753 and Q9Y6N6), mouse laminin (see for example U

[00136] For example, the laminins of the present invention are homologs / orthologs of the laminins of the present invention, such as those of the uniProt accession numbers Q61789, Q61087, and Q61092, chimpanzee laminins (see, e.g., UniProt accession numbers H2QEC7, H2R041, and H2Q0R2), rat laminins (see, e.g., UniProt accession numbers D3ZN05, F1LPI5, and F1LRH4), rabbit laminins (see, e.g., UniProt accession numbers G1SY40 and A0A0B5JSH0), etc. Methods for identifying laminin homologs / orthologs from additional species are known to those of ordinary skill in the art. In some embodiments, the laminin proteins of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any laminin protein described herein or known in the art.

[0225] In some embodiments, the polynucleotides of the present disclosure encode human laminin, such as human laminin subunit alpha-1 (LamA1) polypeptide (see, e.g., UniProt Accession No. P25391), human laminin subunit alpha-2 (Lama2) polypeptide (see, e.g., UniProt Accession No. P24043), human laminin subunit alpha-3 (LamA3) polypeptide (see, e.g., UniProt Accession No. Q16787), human laminin subunit alpha-4 (LamA4) polypeptide (see, e.g., UniProt Accession No. Q16363), human laminin subunit alpha-5 (Lama5) polypeptide (see, e.g., UniProt Accession No. O15230), human laminin subunit alpha-6 (LamA7) polypeptide (see, e.g., UniProt Accession No. Q16616), human laminin subunit alpha-7 (LamA8) polypeptide (see, e.g., UniProt Accession No. Q16617), human laminin subunit alpha-8 (LamA9) polypeptide (see, e.g., UniProt Accession No. Q21426), human laminin subunit alpha-9 (LamA10) polypeptide (see, e.g., UniProt Accession No. Q21427), human laminin subunit alpha-10 (LamA110) polypeptide (see, e.g., UniProt Accession No. Q21426), human laminin subunit alpha-11 (LamA120) polypeptide (see, e.g., UniProt Accession No. Q21426), human laminin subunit alpha-12 (LamA130) polypeptide (see, e.g., UniProt Accession No. Q21426), human laminin subunit alpha-13 (Lam

[0063] The laminin protein subunit β-1 (LamB1) polypeptide (see, e.g., UniProt Accession No. P07942), the laminin protein subunit β-2 (LamB2) polypeptide (see, e.g., UniProt Accession No. P55268), the laminin protein subunit β-3 (LamB3) polypeptide (see, e.g., UniProt Accession No. Q13751), the laminin protein subunit γ-1 (LamC1) polypeptide (see, e.g., UniProt Accession No. P11047), the laminin protein subunit γ-2 (LamC2) polypeptide (see, e.g., UniProt Accession No. Q13753), the laminin protein subunit γ-3 (LamC3) polypeptide (see, e.g., UniProt Accession No. Q9Y6N6), and the like.

[0226] In some embodiments, the polynucleotides of the present disclosure encode human LamA3 polypeptides. In some embodiments, the polynucleotides encoding human LamA3 polypeptides are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 57. In some embodiments, the polynucleotides encoding human LamA3 polypeptides are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 57.

[0227] In some embodiments, the polynucleotide encoding the human LamA3 polypeptide is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 57. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750, at least 3000, at least 3250 but less than 3333 consecutive amino acids of SEQ ID NO: 57.

[0228] In some embodiments, the polynucleotides of the present disclosure encode human LamB3 polypeptides. In some embodiments, the polynucleotides encoding human LamB3 polypeptides are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 58. In some embodiments, the polynucleotides encoding human LamB3 polypeptides are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 58.

[0229] In some embodiments, the polynucleotide encoding the LamB3 polypeptide is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 58. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100 but less than 1172 consecutive amino acids of SEQ ID NO: 58.

[0230] In some embodiments, the polynucleotides of the present disclosure encode human LamC2 polypeptides. In some embodiments, the polynucleotides encoding human LamC2 polypeptides are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 59. In some embodiments, the polynucleotides encoding human LamC2 polypeptides are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 59.

[0231] In some embodiments, the polynucleotide encoding the LamC2 polypeptide is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 59. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100 but less than 1193 consecutive amino acids of SEQ ID NO: 59.

[0232] Neuromodulators

[0233] In some embodiments, the present disclosure relates to one or more polynucleotides encoding full-length neuromodulator proteins or any isoforms or portions thereof. Any neuromodulator protein from any suitable species known in the art may be encoded by the polynucleotides of the present disclosure, including, for example, Clostridium botulinum proteins (see, for example, UniProt accession numbers P0DPI0, Q45894, P0DPI1, P10844, and B1INP5), etc. Methods for identifying homologs / orthologs of neuromodulator proteins from additional species are known to those of ordinary skill in the art. In some embodiments, the neuromodulator proteins of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any neuromodulator protein described herein or known in the art.

[0234] In some embodiments, the polynucleotides of the present disclosure encode Clostridium botulinum neuromodulator proteins.

[0235] In some embodiments, the polynucleotides of the present disclosure encode Clostridium botulinum type A neurotoxin proteins. In some embodiments, the polynucleotides encoding Clostridium botulinum type A neurotoxin proteins are polynucleotides encoding polypeptides comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 60. In some embodiments, the polynucleotides encoding Clostridium botulinum type A neurotoxin proteins are polynucleotides encoding polypeptides comprising the amino acid sequence of SEQ ID NO: 60. In some embodiments, the Clostridium botulinum type A neurotoxin proteins of the present disclosure comprise an alanine to valine mutation at a position corresponding to position 27 of SEQ ID NO: 60.

[0236] In some embodiments, the polynucleotide encoding the Clostridium botulinum type A neurotoxin protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 60. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200 but less than 1296 consecutive amino acids of SEQ ID NO: 60.

[0237] In some embodiments, the polynucleotides of the present disclosure encode Clostridium botulinum type B neurotoxin proteins. In some embodiments, the polynucleotides encoding Clostridium botulinum type B neurotoxin proteins are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of SEQ ID NO: 61. In some embodiments, the polynucleotides encoding Clostridium botulinum type B neurotoxin proteins are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 61.

[0238] In some embodiments, the polynucleotide encoding the Clostridium botulinum type B neurotoxin protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 61. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, at least 1000, at least 1100, at least 1200 but less than 1291 consecutive amino acids of SEQ ID NO: 61.

[0239] Fibrillin

[0240] In some embodiments, the disclosure relates to one or more polynucleotides encoding full-length fibrillin or any subtype or part thereof. Any fibrillin from any suitable species known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, human fibrillin (see, for example, UniProt accession numbers P35555, P35556, and Q75N90), mouse fibrillin (see, for example, UniProt accession numbers Q61554 and Q61555), chimpanzee fibrillin (see, for example, UniProt accession numbers A0A2I3RTE4 and K7CZX0), rat fibrillin (see, for example, UniProt accession numbers G3V9M6 and F1M5Q4), rabbit fibrillin (see, for example, UniProt accession numbers G1SKM2, G1SUS5, and G1T1H4), etc. The method of identifying the fibrillin homologues / orthologues from additional species is known to those of ordinary skill in the art. In some embodiments, the fibrillin proteins of the present disclosure comprise a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any fibrillin protein described herein or known in the art.

[0241] In some embodiments, the polynucleotides of the present disclosure encode human fibrillin.

[0242] In some embodiments, the polynucleotides of the present disclosure encode human fibrillin-1 protein. In some embodiments, the polynucleotides encoding human fibrillin-1 protein are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 62. In some embodiments, the polynucleotides encoding human fibrillin-1 protein are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 62.

[0243] In some embodiments, the polynucleotide encoding human fibrillin-1 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 62. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750 but less than 2871 consecutive amino acids of SEQ ID NO: 62.

[0244] In some embodiments, the polynucleotides of the present disclosure encode human fibrillin-2 protein. In some embodiments, the polynucleotides encoding human fibrillin-2 protein are polynucleotides encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 63. In some embodiments, the polynucleotides encoding human fibrillin-2 protein are polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 63.

[0245] In some embodiments, the polynucleotide encoding human fibrillin-2 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 63. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750 but less than 2912 consecutive amino acids of SEQ ID NO: 63.

[0246] In some embodiments, the polynucleotides of the present disclosure encode human fibrillin-3 protein. In some embodiments, the polynucleotide encoding human fibrillin-3 protein is a polynucleotide encoding a polypeptide comprising an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the sequence of SEQ ID NO: 64. In some embodiments, the polynucleotide encoding human fibrillin-3 protein is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 64.

[0247] In some embodiments, the polynucleotide encoding human fibrillin-3 protein is a polynucleotide encoding an N-terminal truncated form, a C-terminal truncated form, or a fragment of the amino acid sequence of SEQ ID NO: 64. The N-terminal truncated form, the C-terminal truncated form, or a fragment may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 200, at least 300, at least 400, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750 but less than 2809 consecutive amino acids of SEQ ID NO: 64.

[0248] Exemplary cosmetic peptides

[0249] In some embodiments, one or more cosmetic proteins of the present disclosure (e.g., the first cosmetic protein, another cosmetic protein, additional cosmetic protein, and / or the second cosmetic protein) comprise an amino acid sequence comprising at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 15-21 or 53-64. In some embodiments, one or more cosmetic proteins of the present disclosure (e.g., the first cosmetic protein, another cosmetic protein, additional cosmetic protein, and / or the second cosmetic protein) comprise a sequence selected from SEQ ID NOs: 15-21 or 53-64.

[0250] In some embodiments, one or more cosmetic proteins of the present disclosure (e.g., the first cosmetic protein, another cosmetic protein, additional cosmetic protein, and / or the second cosmetic protein) comprise an amino acid sequence comprising at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 15-21, 53-54, or 57-59. In some embodiments, one or more cosmetic proteins of the present disclosure (e.g., the first cosmetic protein, another cosmetic protein, additional cosmetic protein, and / or the second cosmetic protein) comprise a sequence selected from SEQ ID NOs: 15-21, 53-54, or 57-59.

[0251] First polynucleotide

[0252] In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising a first polynucleotide encoding a first polypeptide, wherein the first polypeptide comprises a first cosmetic protein. The first cosmetic protein may be any cosmetic protein described herein or known in the art, including, for example, collagen, fibronectin, elastin, aphrodisiac, vitronectin / vitronectin receptor, laminin, neuromodulators, fibrillin, and the like. In some embodiments, the first cosmetic protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, and the like). In some embodiments, the first cosmetic protein is collagen, elastin, fibronectin, or laminin (e.g., human collagen, elastin, fibronectin, or laminin).

[0253] In some embodiments, the recombinant nucleic acid of the present disclosure comprises one copy of the first polynucleotide. In some embodiments, the recombinant nucleic acid of the present disclosure comprises two or more (e.g., two or more, three or more, four or more, five or more, ten or more, etc.) copies of the first polynucleotide. In some embodiments, the recombinant nucleic acid of the present disclosure comprises two copies of the first polynucleotide.

[0254] In some embodiments, the first cosmetic protein is a first human collagen. The first human collagen can be any human collagen described herein or known in the art. In some embodiments, the first human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, or COL28. In some embodiments, the first human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7, or COL17. In some embodiments, the first human collagen is COL1-1. In some embodiments, the first human collagen is COL1-2. In some embodiments, the first human collagen is COL3. In some embodiments, the first human collagen is COL4-1. In some embodiments, the first human collagen is COL4-2. In some embodiments, the first human collagen is COL6-1. In some embodiments, the first human collagen is COL7. In some embodiments, the first human collagen is not COL7. In some embodiments, the first human collagen is COL17.

[0255] In some embodiments, the first polypeptide consists essentially of the first cosmetic protein. In some embodiments, the first polypeptide consists of the first cosmetic protein. In some embodiments, the first polypeptide is the first cosmetic protein.

[0256] chimeric polygon

[0257] In some embodiments, the first polypeptide is a chimeric polypeptide comprising a first beauty protein. In some embodiments, the first polypeptide is a chimeric polypeptide comprising a first beauty protein and another beauty protein. In some embodiments, the chimeric polypeptide comprises a linker polypeptide connecting the first beauty protein and another beauty protein. In some embodiments, the chimeric polypeptide comprises a first beauty protein-linker polypeptide-another beauty protein from the n-terminus to the c-terminus. The first beauty protein and / or another beauty protein can be any beauty protein described herein or known in the art, including, for example, collagen, fibronectin, elastin, aphrodisiac, vitronectin / vitronectin receptor, laminin, neuromodulator, fibrillin, etc. In some embodiments, the first beauty protein and / or another beauty protein are structural extracellular matrix proteins (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the first beauty protein and / or another beauty protein are collagen, elastin, fibronectin or laminin (e.g., human collagen, elastin, fibronectin or laminin). In some embodiments, the first beauty protein and another beauty protein are the same. In some embodiments, the first cosmetic protein and the another cosmetic protein are different.

[0258] In some embodiments, the linker polypeptide is a cleavable linker polypeptide. Any cleavable linker polypeptide known in the art can be used in the chimeric polypeptides of the present disclosure, including, for example, T2A linkers, P2A linkers, E2A linkers, and F2A linkers, among others. In some embodiments, the linker polypeptide is a T2A linker polypeptide. An exemplary nucleic acid sequence encoding a T2A linker polypeptide is provided as SEQ ID NO: 24. An exemplary amino acid sequence of a T2A linker polypeptide is provided as SEQ ID NO: 28. In some embodiments, the linker polypeptide is a P2A linker polypeptide. An exemplary nucleic acid sequence encoding a P2A linker polypeptide is provided as SEQ ID NO: 25. An exemplary amino acid sequence of a P2A linker polypeptide is provided as SEQ ID NO: 29. In some embodiments, the linker polypeptide is an E2A linker polypeptide. An exemplary nucleic acid sequence encoding an E2A linker polypeptide is provided as SEQ ID NO: 26. An exemplary amino acid sequence of an E2A linker polypeptide is provided as SEQ ID NO: 30. In some embodiments, the linker polypeptide is a F2A linker polypeptide. An exemplary nucleic acid sequence encoding an F2A linker polypeptide is provided as SEQ ID NO: 27. An exemplary amino acid sequence of a F2A linker polypeptide is provided as SEQ ID NO:31.

[0259] In some embodiments, the linker polypeptide comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 28-31. In some embodiments, the linker polypeptide comprises a sequence selected from SEQ ID NOs: 28-31.

[0260] In some embodiments, the first cosmetic protein is a first collagen (e.g., a first human collagen), and the other cosmetic protein is another collagen (e.g., another human collagen). An exemplary nucleic acid sequence encoding a chimeric polypeptide comprising a first human collagen, a linker polypeptide, and another human collagen is provided as SEQ ID NO: 32.

[0261] In some embodiments, the first cosmetic protein is a first human collagen and the other cosmetic protein is another human collagen. The other human collagen can be any human collagen described herein or known in the art. In some embodiments, the other human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, or COL28. In some embodiments, the other human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7, or COL17. In some embodiments, the other human collagen is COL1-1. In some embodiments, the other human collagen is COL1-2. In some embodiments, the other human collagen is COL3. In some embodiments, the other human collagen is COL4-1. In some embodiments, the other human collagen is COL4-2. In some embodiments, the other human collagen is COL6-1. In some embodiments, the other human collagen is COL7. In some embodiments, the other human collagen is not COL7. In some embodiments, the other human collagen is COL17. In some embodiments, the first human collagen and the other human collagen are the same. In some embodiments, the first human collagen and the other human collagen are different.

[0262] In some embodiments, the first human collagen is COL1-1 and the other human collagen is selected from COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL1-1 and the other human collagen is COL1-2. In some embodiments, the first human collagen is COL1-1 and the other human collagen is COL3.

[0263] In some embodiments, the first human collagen is COL1-2 and the other human collagen is COL1-1, COL3, COL4-1, COL4-2, COL6-1, COL7, or COL17. In some embodiments, the first human collagen is COL1-2 and the other human collagen is COL1-1.

[0264] In some embodiments, the first human collagen is COL3 and the other human collagen is selected from COL1-1, COL1-2, COL4-1, COL4-2, COL6-1, COL7 or COL17.

[0265] In some embodiments, the first human collagen is COL4-1 and the other human collagen is COL1-1, COL1-2, COL3, COL4-2, COL6-1, COL7 or COL 17. In some embodiments, the first human collagen is COL4-1 and the other human collagen is COL4-2.

[0266] In some embodiments, the first human collagen is COL6-1 and the other human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL7 or COL17.

[0267] In some embodiments, the first human collagen is COL7 and the other human collagen is COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, or COL17.

[0268] In some embodiments, the first human collagen is COL17 and the other human collagen is COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, or COL7.

[0269] In some embodiments, the first cosmetic protein is a first laminin (e.g., a first human laminin), and the other cosmetic protein is another laminin (e.g., another human laminin). In some embodiments, the first cosmetic protein is a first human laminin, and the other cosmetic protein is another human laminin. The other human laminin may be any human laminin described herein or known in the art. In some embodiments, the first human laminin is a human LamA3 polypeptide, and the other human laminin is a human LamB3 polypeptide. In some embodiments, the first human laminin is a human LamA3 polypeptide, and the other human laminin is a human LamC2 polypeptide. In some embodiments, the first human laminin is a human LamB3 polypeptide, and the other human laminin is a human LamC2 polypeptide.

[0270] In some embodiments, the first polynucleotide encodes a monocistronic mRNA. In some embodiments, the monocistronic mRNA comprises an open reading frame (ORF) encoding a first polypeptide.

[0271] In some embodiments, the first polynucleotide encodes a polycistronic mRNA.In some embodiments, the polycistronic mRNA comprises an open reading frame (ORF) encoding a first polypeptide.

[0272] Polycistronic mRNA

[0273] In some embodiments, the first polynucleotide encodes a polycistronic mRNA. In some embodiments, the polycistronic mRNA comprises an open reading frame (ORF) encoding a first polypeptide. In some embodiments, the first polynucleotide encodes a polycistronic mRNA, and the polycistronic mRNA comprises: 1) a first open reading frame (ORF) encoding a first polypeptide, and 2) a second open reading frame (ORF) encoding an additional beauty protein. In some embodiments, the polycistronic mRNA also comprises an internal ribosome entry site (IRES) separating the first ORF and the second ORF. In some embodiments, the polycistronic mRNA comprises a first ORF-IRES-encoding a second ORF of an additional beauty protein from 5' to 3' encoding a first polypeptide. The first polypeptide may be any first polypeptide described herein. The additional beauty protein may be any beauty protein described herein or known in the art, including, for example, collagen, fibronectin, elastin, aphrodisiac, vitronectin / vitronectin receptor, laminin, neuromodulators, fibrillin, etc. In some embodiments, the additional cosmetic protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the additional cosmetic protein is collagen, elastin, fibronectin, or laminin (e.g., human collagen, elastin, fibronectin, or laminin).

[0274] Any suitable IRES known in the art can be used in the polycistronic mRNA of the present disclosure, including, for example, viral-derived IRES (e.g., IRES derived from poliovirus, rhinovirus, encephalomyocarditis virus (EMCV), foot-and-mouth disease virus, hepatitis C virus, classical swine fever virus, rous sarcoma virus, human immunodeficiency virus, cricket paralysis virus, Kaposi's sarcoma-associated herpes virus, etc.), cellular mRNA-derived IRES (e.g., IRES derived from growth factor mRNAs such as fibroblast growth factor 2, platelet-derived growth factor B, and vascular endothelial growth factor; IRES derived from transcription factor mRNAs such as antennapedia, ultrabithorax, and NF-κB repressor; IRES derived from oncogene mRNAs such as c-myc, pim-1, and protein kinase p58) PITSLRE et al.), synthetic IRES (e.g., CP148 IRES), and other IRESs (see, e.g., Mokrej s et al. (2007) A Bioinformatical Approach to the Analysis of Viral and Cellular Internal Ribosome Entry Sites. Columbus F, ed. New Messenger RNA Research Communications. Hauppauge, NY: Nova Science Publishers; pp. 133-166). In some embodiments, the IRES is a CP148 IRES. An exemplary nucleic acid sequence encoding a CP148 IRES is provided as SEQ ID NO: 22. In some embodiments, the IRES is an EMCV IRES. An exemplary nucleic acid sequence encoding an EMCV IRES is provided as SEQ ID NO: 23.

[0275] In some embodiments, the nucleic acid sequence encoding an IRES comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleic acid sequence selected from SEQ ID NO: 22 or SEQ ID NO: 23. In some embodiments, the nucleic acid sequence encoding an IRES comprises the sequence of SEQ ID NO: 22 or SEQ ID NO: 23.

[0276] In some embodiments, the first polypeptide is a first collagen (e.g., a first human collagen), and the additional cosmetic protein is an additional collagen (e.g., an additional human collagen). An exemplary nucleic acid encoding a polycistronic mRNA comprising a first ORF, an IRES, and a second ORF is provided as SEQ ID NO: 33 or SEQ ID NO: 34. The additional human collagen can be any of the human collagens described herein. In some embodiments, the additional human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27, or COL28. In some embodiments, the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7, or COL17. In some embodiments, the additional human collagen is COL1-1. In some embodiments, the additional human collagen is COL1-2. In some embodiments, the additional human collagen is COL3. In some embodiments, the additional human collagen is COL4-1. In some embodiments, the additional human collagen is COL4-2. In some embodiments, the additional human collagen is COL6-1. In some embodiments, the additional human collagen is COL7. In some embodiments, the additional human collagen is not COL7. In some embodiments, the additional human collagen is COL17. In some embodiments, the first human collagen and the additional human collagen are the same. In some embodiments, the first human collagen and the additional human collagen are different.

[0277] In some embodiments, the first human collagen is COL1-1 and the additional human collagen is selected from COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL1-1 and the additional human collagen is COL1-2. In some embodiments, the first human collagen is COL1-1 and the additional human collagen is COL3.

[0278] In some embodiments, the first human collagen is COL1-2 and the additional human collagen is selected from COL1-1, COL3, COL4-1, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL1-2 and the additional human collagen is COL1-1.

[0279] In some embodiments, the first human collagen is COL3 and the additional human collagen is selected from COL1-1, COL1-2, COL4-1, COL4-2, COL6-1, COL7 or COL17.

[0280] In some embodiments, the first human collagen is COL4-1 and the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL4-1 and the additional human collagen is COL4-2.

[0281] In some embodiments, the first human collagen is COL6-1 and the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL7 or COL17.

[0282] In some embodiments, the first human collagen is COL7 and the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, or COL17.

[0283] In some embodiments, the first human collagen is COL17 and the additional human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, or COL7.

[0284] In some embodiments, the first polypeptide is a first collagen (eg, a first human collagen), and the additional cosmetic protein is an additional collagen (eg, an additional human collagen).

[0285] In some embodiments, the first polypeptide is a first laminin (e.g., a first human laminin), and the additional beauty protein is an additional laminin (e.g., an additional human laminin). In some embodiments, the first polypeptide is a first human laminin, and the additional beauty protein is an additional human laminin. The additional human laminin may be any human laminin described herein or known in the art. In some embodiments, the first human laminin is a human LamA3 polypeptide, and the additional human laminin is a human LamB3 polypeptide. In some embodiments, the first human laminin is a human LamA3 polypeptide, and the additional human laminin is a human LamC2 polypeptide. In some embodiments, the first human laminin is a human LamB3 polypeptide, and the additional human laminin is a human LamC2 polypeptide.

[0286] The second polynucleotide

[0287] In some embodiments, the present disclosure relates to a recombinant nucleic acid further comprising a second polynucleotide encoding a second beauty protein. The second beauty protein may be any beauty protein described herein or known in the art, including, for example, collagen, fibronectin, elastin, aphrodisiac, vitronectin / vitronectin receptor, laminin, neuromodulator, fibrillin, etc. In some embodiments, the second beauty protein is a structural extracellular matrix protein (e.g., collagen, elastin, fibronectin, laminin, fibrillin, etc.). In some embodiments, the second beauty protein is collagen, elastin, fibronectin or laminin (e.g., human collagen, elastin, fibronectin or laminin). In some embodiments, the first beauty protein and the second beauty protein are the same. In some embodiments, the first beauty protein and the second beauty protein are different. In some embodiments, the recombinant nucleic acid comprises a copy of the second polynucleotide. In some embodiments, the recombinant nucleic acid comprises two or more (e.g., two or more, three or more, four or more, five or more, ten or more, etc.) copies of the second polynucleotide. In some embodiments, the recombinant nucleic acid comprises two copies of the second polynucleotide.

[0288] In some embodiments, the second cosmetic protein is collagen. In some embodiments, the second cosmetic protein is a second human collagen. The second human collagen can be any human collagen described herein. In some embodiments, the second human collagen is selected from COL1-1, COL1-2, COL2, COL3, COL4-1, COL4-2, COL4-3, COL4-4, COL4-5, COL4-6, COL5-1, COL5-2, COL5-3, COL6-1, COL6-2, COL6-3, COL6-4, COL6-5, COL6-6, COL7 , COL8, COL9-1, COL9-2, COL9-3, COL10, COL11-1, COL11-2, COL12, COL13, COL14, COL15, COL16, COL17, COL18, COL19, COL20, COL21, COL22, COL23, COL24, COL25, COL26, COL27 or COL28. In some embodiments, the second human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL6-1, COL7 or COL17. In some embodiments, the second human collagen is COL1-1. In some embodiments, the second human collagen is COL1-2. In some embodiments, the second human collagen is COL3. In some embodiments, the second human collagen is COL4-1. In some embodiments, the second human collagen is COL4-2. In some embodiments, the second human collagen is COL6-1. In some embodiments, the second human collagen is COL7. In some embodiments, the second human collagen is not COL7. In some embodiments, the second human collagen is COL17.

[0289] In some embodiments, the first polynucleotide encodes a first collagen protein, and the second polynucleotide encodes a second collagen protein. In some embodiments, the first polynucleotide encodes a first human collagen protein, and the second polynucleotide encodes a second human collagen protein. In some embodiments, the first human collagen protein (encoded by the first polynucleotide) and the second human collagen protein (encoded by the second polynucleotide) are the same. In some embodiments, the first human collagen protein (encoded by the first polynucleotide) and the second human collagen protein (encoded by the second polynucleotide) are different.

[0290] In some embodiments, the first human collagen is COL1-1, and the second human collagen is selected from COL1-2, COL3, COL4-1, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL1-1, and the second human collagen is COL1-2. In some embodiments, the first human collagen is COL1-1, and the second human collagen is COL3.

[0291] In some embodiments, the first human collagen is COL1-2 and the second human collagen is selected from COL1-1, COL3, COL4-1, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL1-2 and the second human collagen is COL1-1.

[0292] In some embodiments, the first human collagen is COL3 and the second human collagen is selected from COL1-1, COL1-2, COL4-1, COL4-2, COL6-1, COL7 or COL17.

[0293] In some embodiments, the first human collagen is COL4-1 and the second human collagen is selected from COL1-2, COL1-2, COL3, COL4-2, COL6-1, COL7 or COL17. In some embodiments, the first human collagen is COL4-1 and the second human collagen is COL4-2.

[0294] In some embodiments, the first human collagen is COL6-1 and the second human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL7 or COL17.

[0295] In some embodiments, the first human collagen is COL7 and the second human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, or COL17.

[0296] In some embodiments, the first human collagen is COL17 and the second human collagen is selected from COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1, or COL7.

[0297] In some embodiments, the first polynucleotide encodes a first laminin (e.g., a first human laminin), and the second polynucleotide encodes a second laminin (e.g., a second human laminin). In some embodiments, the first polynucleotide encodes a first human laminin polypeptide, and the second polynucleotide encodes a second human laminin. The second human laminin may be any human laminin described herein or known in the art. In some embodiments, the first human laminin is a human LamA3 polypeptide, and the second human laminin is a human LamB3 polypeptide. In some embodiments, the first human laminin is a human LamA3 polypeptide, and the second human laminin is a human LamC2 polypeptide. In some embodiments, the first human laminin is a human LamB3 polypeptide, and the second human laminin is a human LamC2 polypeptide.

[0298] Recombinant nucleic acid

[0299] In some embodiments, the disclosure relates to a recombinant nucleic acid comprising any one or more of the polynucleotides described herein. In some embodiments, the recombinant nucleic acid comprises a copy of the first polynucleotide. In some embodiments, the recombinant nucleic acid comprises two copies of the first polynucleotide. In some embodiments, the recombinant nucleic acid comprises a copy of the first polynucleotide and a copy of the second polynucleotide. In some embodiments, the recombinant nucleic acid comprises a copy of the first polynucleotide and two copies of the second polynucleotide. In some embodiments, the recombinant nucleic acid comprises two copies of the first polynucleotide and one copy of the second polynucleotide. In some embodiments, the recombinant nucleic acid comprises two copies of the first polynucleotide and two copies of the second polynucleotide.

[0300] In some embodiments, the recombinant nucleic acid is a vector (e.g., an expression vector, a display vector, etc.). In some embodiments, the vector is a DNA vector or an RNA vector. Generally, a vector suitable for maintaining, propagating, and / or expressing polynucleotides in a subject to produce one or more polypeptides can be used. Examples of suitable vectors may include, for example, plasmids, cosmids, episomes, transposons, and viral vectors (e.g., adenovirus vectors, adeno-associated virus vectors, vaccinia virus vectors, Sindbis virus vectors, measles vectors, herpes virus vectors, lentiviral vectors, retroviral vectors, etc.). In some embodiments, the vector is a herpes virus vector. In some embodiments, the vector can replicate autonomously in a host cell. In some embodiments, the vector cannot replicate autonomously in a host cell. In some embodiments, the vector can be integrated into a host DNA. In some embodiments, the vector cannot be integrated into a host DNA (e.g., an episomal type). The method for preparing a vector containing one or more target polynucleotides is well known to those of ordinary skill in the art, including, for example, by chemical synthesis, or by artificially manipulating a separated segment of a nucleic acid (e.g., by genetic engineering techniques).

[0301] In some embodiments, the recombinant nucleic acid of the present disclosure is a herpes simplex virus (HSV) amplicon. Herpes simplex virus amplicon, including structural features and methods for preparing the herpes simplex virus amplicon, is generally known to those of ordinary skill in the art (see, for example, de Silva S. and Bowers W. "Herpes Vires Amplicon Vectors". Viruses 2009, 1, 594-629). In some embodiments, the herpes simplex virus amplicon is an HSV-1 amplicon. In some embodiments, the herpes simplex virus amplicon is an HSV-1 hybrid amplicon. Examples of HSV-1 hybrid amplicons may include, but are not limited to, HSV / AAV hybrid amplicon, HSV / EBV hybrid amplicon, HSV / EBV / RV hybrid amplicon, and / or HSV / Sleeping Beauty hybrid amplicon. In some embodiments, the amplicon is an HSV / AAV hybrid amplicon. In some embodiments, the amplicon is an HSV / Sleeping Beauty hybrid amplicon.

[0302] In some embodiments, the recombinant nucleic acid of the present disclosure is a recombinant herpesvirus genome.The recombinant herpesvirus genome can be the recombinant genome of any member of the Herpesviridae from DNA viruses as known in the art, including for example recombinant herpes simplex virus genome, recombinant varicella zoster virus genome, recombinant human cytomegalovirus genome, recombinant herpesvirus 6A genome, recombinant herpesvirus 6B genome, recombinant herpesvirus 7 genome, recombinant Kaposi's sarcoma-associated herpesvirus genome and any combination or any derivative thereof.In some embodiments, the recombinant herpesvirus genome comprises one or more (for example one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) inactivation mutations.In some embodiments, one or more inactivation mutations are in one or more (for example one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) herpesvirus genes. In some embodiments, the recombinant herpesvirus genome is attenuated (e.g., compared to the corresponding wild-type herpesvirus genome). In some embodiments, the recombinant herpesvirus genome is replication-competent. In some embodiments, the recombinant herpesvirus genome is replication-defective.

[0303] In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus (HSV) genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivative thereof. In some embodiments, the recombinant herpes simplex virus genome is a recombinant HSV-1 genome. In some embodiments, the recombinant herpes simplex virus genome is replication-competent. In some embodiments, the recombinant herpes simplex virus genome is replication-defective. In some embodiments, the recombinant herpes simplex virus genome comprises one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) inactivation mutations. In some embodiments, one or more inactivation mutations are in one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) herpes simplex virus genes. As used herein, "inactivating mutation" may refer to any mutation that causes a gene or regulator product (RNA or protein) to have a reduced, undetectable or eliminated quantity and / or function (e.g., compared to the corresponding sequence lacking an inactivating mutation). The example of an inactivating mutation may include, but is not limited to, deletions, insertions, point mutations and rearrangements in the transcriptional control sequence (promoter, enhancer, insulator, etc.) and / or coding sequence of a given gene or regulator. Any suitable method for measuring the quantity of a gene or regulator product as known in the art may be used, including, for example, qPCR, Northern blot, RNA sequencing, protein blot, ELISA, etc.

[0304] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in at least one, at least two, at least three, at least four, at least five, at least six, at least seven, or all eight of the infected cell protein (or infected cell polypeptide) (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41, and / or UL55 herpes simplex virus genes. In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation in the ICP34.5 and / or ICP47 herpes simplex virus genes (e.g., to avoid the production of immunostimulatory viruses). In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation in the ICP34.5 herpes simplex virus gene (one or two copies). In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation in the ICP47 herpes simplex virus gene. In some embodiments, the recombinant herpes simplex virus genome does not comprise inactivating mutations in the ICP34.5 (one or two copies) and ICP47 herpes simplex virus genes. In some embodiments, the recombinant herpes simplex virus genome is not oncolytic.

[0305] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), and also comprises a priming mutation in ICP4 (one or two copies), ICP22, ICP27, ICP47, UL41 and / or UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), and comprises an inactivating mutation in the ICP4 gene (one or two copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), and comprises an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), and comprises an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), an inactivating mutation in the ICP4 gene (one or two copies), and an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), an inactivating mutation in the ICP4 gene (one or two copies), and an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), an inactivating mutation in the ICP22 gene, and an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICPO gene (one or two copies), an inactivating mutation in the ICP4 gene (one or two copies), an inactivating mutation in the ICP22 gene, and an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of ICP0 (one or two copies), ICP4 (one or two copies), ICP22 and / or UL41 genes. In some embodiments, the recombinant herpes simplex virus genome also comprises an inactivating mutation in ICP27, ICP47 and / or UL55 genes.

[0306] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or two copies). In some embodiments, the recombinant compound herpes (herpes complex) virus genome comprises an inactivating mutation in ICP4 (one or two copies), and also comprises an inactivating mutation in ICP0 (one or two copies), ICP22, ICP27, ICP47, UL41 and / or UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or two copies), and comprises an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or two copies), and comprises an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or two copies), comprises an inactivating mutation in the ICP22 gene, and comprises an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of ICP4 (one or two copies), ICP22 and / or UL41 genes. In some embodiments, the recombinant herpes simplex virus genome also comprises an inactivating mutation in ICP0, ICP27, ICP47 and / or UL55 genes.

[0307] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene, and also comprises an inactivating mutation in ICPO (one or two copies), ICP4 (one or two copies), ICP27, ICP47, UL41 and / or UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene, and comprises an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is the deletion of the coding sequence of ICP22 and / or UL41 genes. In some embodiments, the recombinant herpes simplex virus genome also comprises an inactivating mutation in ICPO (one or two copies), ICP4 (one or two copies), ICP27, ICP47 and / or UL55 genes.

[0308] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene, and also comprises an inactivating mutation in ICPO (one or two copies), ICP4 (one or two copies), ICP22, ICP47, UL41 and / or UL55 genes. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP27 gene.

[0309] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP47 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP47 gene and also comprises an inactivating mutation in ICPO (one or two copies), ICP4 (one or two copies), ICP22, ICP27, UL41 and / or UL55 genes. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP47 gene.

[0310] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL41 gene, and also comprises an inactivating mutation in ICP0 (one or two copies), ICP4 (one or two copies), ICP22, ICP27, ICP47 and / or UL55 genes. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the UL41 gene.

[0311] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene and also comprises an inactivating mutation in ICP0 (one or two copies), ICP4 (one or two copies), ICP22, ICP27, ICP47 and / or UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the UL55 gene.

[0312] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation (e.g., a deletion of the internal repeat (junction) region) in an internal repeat long (IRL) region and an internal repeat short (IRS) region. In some embodiments, the inactivation (e.g., deletion) of the junction region eliminates one copy of each of the ICP4 gene and the ICPO gene. In some embodiments, the inactivation (e.g., deletion) of the junction region further inactivates (e.g., deletes) the promoters of the ICP22 and ICP47 genes. If desired, the expression of one or both of these genes can be restored by inserting an immediate early promoter into the recombinant herpes simplex virus genome (see, e.g., Hill et al. (1995). Nature 375 (6530): 411-415; Goldsmith et al. (1998). J Exp Med 187 (3): 341-348). Without wishing to be bound by theory, it is believed that inactivating (eg, deleting) the junction region may promote stability of the recombinant herpes simplex virus genome and / or allow the recombinant herpes simplex virus genome to accommodate more and / or larger transgenes.

[0313] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in ICP4 (one or two copies), ICP22 and ICP27 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in ICP4 (one or two copies), ICP27 and UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in ICP4 (one or two copies), ICP22, ICP27, ICP47 and UL55 genes. In some embodiments, the inactivating mutation in ICP4 (one or two copies), ICP27 and / or UL55 genes is a deletion of the coding sequence of ICP4 (one or two copies), ICP27 and / or UL55 genes. In some embodiments, the inactivating mutation in ICP22 and ICP47 genes is a deletion in the promoter region of ICP22 and ICP47 genes (e.g., ICP22 and ICP47 coding sequences are complete, but do not have transcriptional activity). In some embodiments, the recombinant herpes simplex virus genome comprises a deletion in the coding sequence of ICP4 (one or two copies), ICP27 and UL55 genes, and a deletion in the promoter region of ICP22 and ICP47 genes. In some embodiments, the recombinant herpes simplex virus genome also comprises an inactivating mutation in ICP0 and / or UL41 genes.

[0314] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 (one or two copies) gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 (one or two copies) and ICP4 (one or two copies) genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 (one or two copies), ICP4 (one or two copies) and ICP22 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 (one or two copies), ICP4 (one or two copies), ICP22 and ICP27 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 (one or two copies), ICP4 (one or two copies), ICP22, ICP27 and UL55 genes. In some embodiments, the inactivating mutation in ICP0 (one or two copies), ICP4 (one or two copies), ICP22, ICP27 and / or UL55 gene comprises a deletion of the coding sequence of ICP0, ICP4 (one or two copies), ICP22, ICP27 and / or UL55 gene. In some embodiments, the recombinant herpes simplex virus genome also comprises an inactivating mutation in ICP47 and / or UL41 gene.

[0315] In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure in one, two, three, four, five, six, seven or more viral loci. Examples of suitable viral loci may include but are not limited to ICP0 (one or two copies), ICP4 (one or two copies), ICP22, ICP27, ICP47, tk, UL41 and UL55 herpes simplex virus loci. In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure in one or two of the viral ICP4 loci (e.g., the recombinant virus carries the first polynucleotide encoding the first human collagen in one or two of the ICP4 loci; the recombinant virus carries the second polynucleotide encoding the second human collagen in one or two of the ICP4 loci; etc.). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure in the viral ICP22 locus (e.g., the recombinant virus carries the first polynucleotide encoding the first human collagen in the ICP22 locus; the recombinant virus carries the second polynucleotide encoding the second human collagen in the ICP22 locus; etc.). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the viral UL41 locus (e.g., the recombinant virus carries a first polynucleotide encoding a first human collagen in the UL41 locus; the recombinant virus carries a second polynucleotide encoding a second human collagen in the UL41 locus; etc.). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or both of the viral ICP4 loci, and comprises one or more polynucleotides of the present disclosure within the viral ICP22 locus (e.g., the recombinant virus carries a first polynucleotide encoding a first human collagen in one or both of the ICP4 loci, and carries a second polynucleotide encoding a second human collagen in the ICP22 locus; the recombinant virus carries a second polynucleotide encoding a second human collagen in one or both of the ICP4 loci, and carries a first polynucleotide encoding a first human collagen in the ICP22 locus; etc.). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or both of the viral ICP4 loci, and comprises one or more polynucleotides of the present disclosure within the viral UL41 locus (e.g., the recombinant virus carries a first polynucleotide encoding a first human collagen in one or both of the ICP4 loci, and carries a second polynucleotide encoding a second human collagen in the UL41 locus; the recombinant virus carries a second polynucleotide encoding a second human collagen in one or both of the ICP4 loci, and carries a first polynucleotide encoding a first human collagen in the UL41 locus; etc.).In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure in one or both of the viral ICP4 loci, one or more polynucleotides of the present disclosure in the viral ICP22 locus, and one or more polynucleotides of the present disclosure in the viral UL41 locus (e.g., the recombinant virus carries a first polynucleotide encoding a first human collagen in one or both of the ICP4 loci, and carries a second polynucleotide encoding a second human collagen in both the ICP22 and UL41 loci; the recombinant virus carries a second polynucleotide encoding a second human collagen in one or both of the ICP4 loci, and carries a first polynucleotide encoding a first human collagen in both the ICP22 and UL41 loci; etc.).

[0316] In some embodiments, the recombinant herpes virus genome (e.g., a recombinant herpes simplex virus genome) has been engineered to reduce or eliminate the expression of one or more toxic herpes simplex genes (such as one or two copies of the HSV ICP0 gene, one or two copies of the HSV ICP4 gene, the ICP22 gene, and / or the UL41 gene). In some embodiments, the recombinant herpes virus genome (e.g., a recombinant herpes simplex virus genome) has been engineered to reduce the cytotoxicity of the recombinant genome (e.g., when introduced into a target cell) compared to the corresponding wild-type herpes virus genome (e.g., a wild-type herpes simplex virus genome). In some embodiments, the cytotoxicity of a recombinant viral genome (e.g., a recombinant herpes simplex virus genome) is reduced (e.g., in human keratinocytes and / or fibroblasts) by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100%, at least about 101%, at least about 102%, at least about 103%, at least about 104%, at least about 105%, at least about 106%, at least about 107%, at least about 108%, at least about 109%, at least about 110%, at least about 111%, at least about 112%, at least about 113%, at least about 114%, at least about 115%, About 85%, at least about 90%, at least about 95% or at least about 99% (e.g., measuring the relative cytotoxicity of the recombinant ΔICP4 (one or two copies) herpes simplex virus genome relative to the wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); measuring the relative cytotoxicity of the recombinant ΔICP4 (one or two copies) / ΔICP22 herpes simplex virus genome relative to the wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); etc.). In some embodiments, the cytotoxicity of a recombinant herpes virus genome (e.g., a recombinant herpes simplex virus genome) (e.g., in human keratinocytes and / or fibroblasts) is reduced by at least about 1.5, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 15, at least about 120, at least about 125, at least about 150, at least about 175, at least about 100, at least about 1000, at least about 1500, at least about 10 ... 1 / 250, at least about 1 / 500, at least about 1 / 750, at least about 1 / 1000, or more (e.g., measuring the relative cytotoxicity of the recombinant ΔICP4 (one or two copies) herpes simplex virus genome relative to the wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); measuring the relative cytotoxicity of the recombinant ΔICP4 (one or two copies) / ΔICP22 herpes simplex virus genome relative to the wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); etc.).Methods for measuring cytotoxicity are known to those of ordinary skill in the art and include, for example, by using vital dyes (formazan). dyes), protease biomarkers, MTT assay (or assays using related tetrazolium salts such as XTT, MTS, water-soluble tetrazolium salts, etc.), measuring ATP content, etc.

[0317] In some embodiments, compared to corresponding wild-type herpesvirus genome (for example wild-type herpes simplex virus genome), recombinant herpesvirus genome (for example recombinant herpes simplex virus genome) has been engineered to make after target cell is exposed to recombinant genome, the impact of recombinant herpesvirus genome on host cell proliferation is reduced. In some embodiments, target cell is human cell. In some embodiments, target cell is the cell of epidermis and / or dermis. In some embodiments, target cell is keratinocyte and / or fibroblast. In some embodiments, host cell proliferation (e.g., human keratinocytes and / or fibroblasts) after exposure to the recombinant genome is accelerated by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%. about 99% less (e.g., in the case of human keratinocytes or fibroblasts (primary cells or cell lines), the relative cell proliferation after exposure to the recombinant ΔICP4 (one or two copies) herpes simplex virus genome is measured relative to the cell proliferation after exposure to the wild-type herpes simplex virus genome; in the case of human keratinocytes or fibroblasts (primary cells or cell lines), the relative cell proliferation after exposure to the recombinant ΔICP4 (one or two copies) / ΔICP22 herpes simplex virus genome is measured relative to the cell proliferation after exposure to the wild-type herpes simplex virus genome; etc.).In some embodiments, host cell proliferation (e.g., human keratinocytes and / or fibroblasts) after exposure to the recombinant genome is accelerated by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 50 times, at least about 75 times, at least about 100 times, at least about 250 times, at least about 500 times, at least about 750 times, or ... 1000 times (e.g., in the case of human keratinocytes or fibroblasts (primary cells or cell lines), relative to the cell proliferation after exposure to the wild-type herpes simplex virus genome, the relative cell proliferation after exposure to the recombinant ΔICP4 (one or two copies) herpes simplex virus genome is measured; in the case of human keratinocytes or fibroblasts (primary cells or cell lines), relative to the cell proliferation after exposure to the wild-type herpes simplex virus genome, relative cell proliferation after exposure to the recombinant ΔICP4 (one or two copies) / ΔICP22 herpes simplex virus genome is measured; etc.). Methods for measuring cell proliferation are known to those of ordinary skill in the art, including, for example, by using Ki67 cell proliferation assays, BrdU cell proliferation assays, etc.

[0318] A vector (e.g., a herpes virus vector) may include one or more polynucleotides of the present disclosure in a form suitable for expressing the polynucleotide in a host cell. A vector may include one or more regulatory sequences (e.g., as described above) operably linked to the polynucleotide to be expressed.

[0319] In some embodiments, the recombinant nucleic acid (e.g., recombinant herpes simplex virus genome) of the present disclosure comprises one or more of the polynucleotides described herein, and the one or more are inserted into the recombinant nucleic acid in any orientation. If the recombinant nucleic acid comprises two or more polynucleotides (e.g., two or more, three or more, etc.) described herein, the polynucleotides can be inserted in the same orientation or opposite orientation to each other. Without wishing to be bound by theory, incorporating two polynucleotides (e.g., two transgenics) into the recombinant nucleic acid (e.g., vector) in an antisense orientation can help avoid read-through and ensure the appropriate expression of each polynucleotide.

[0320] IV. Viruses

[0321] Some aspects of the present disclosure relate to viruses comprising any one of the polynucleotides and / or recombinant nucleic acids described herein. In some embodiments, the virus can infect one or more target cells of a subject (e.g., a human subject). In some embodiments, the virus is suitable for delivering polynucleotides and / or recombinant nucleic acids to one or more target cells of a subject (e.g., a human subject). In some embodiments, one or more target cells are one or more human cells. In some embodiments, one or more target cells are one or more cells of the skin (e.g., one or more cells of epidermis, dermis, and / or subcutaneous tissue). In some embodiments, one or more cells are selected from keratinocytes, melanocytes, Langerhans cells (Langerhans cell), Merkel cells (Merkel cell), mast cells, fibroblasts, and / or adipocytes. In some embodiments, one or more cells are keratinocytes. In some embodiments, one or more cells are present in stratum corneum, stratum granulosum, stratum spinosum, stratum basale, and / or basal membrane. In some embodiments, one or more target cells are one or more epidermal cells.

[0322] Any suitable virus known in the art can be used, including, for example, adenovirus, adeno-associated virus, retrovirus, slow virus, Sendai virus, herpes virus (e.g., herpes simplex virus), vaccinia virus and / or any hybrid virus thereof. In some embodiments, the virus is attenuated. In some embodiments, the virus is replication-defective. In some embodiments, the virus is replication-competent. In some embodiments, the virus has been modified to change its tissue tropism relative to the tissue tropism of the unmodified wild-type virus. In some embodiments, the virus has reduced cytotoxicity compared to the corresponding wild-type virus. Methods for producing viruses comprising recombinant nucleic acids are well known to those of ordinary skill in the art.

[0323] In some embodiments, virus is a member of the Herpesviridae family of DNA virus, including for example herpes simplex virus, varicella zoster virus, human cytomegalovirus, herpesvirus 6A, herpesvirus 6B, herpesvirus 7 and Kaposi's sarcoma associated herpesvirus etc. In some embodiments, herpesvirus is attenuated. In some embodiments, herpesvirus is replication defective. In some embodiments, herpesvirus is replication competent. In some embodiments, herpesvirus has reduced cytotoxicity compared to corresponding wild-type herpesvirus. In some embodiments, herpesvirus is not oncolytic.

[0324] In some embodiments, the virus is herpes simplex virus. The herpes simplex virus comprising a recombinant nucleic acid can be produced by a method disclosed, for example, in WO2015 / 009952 and / or WO2017 / 176336. In some embodiments, the herpes simplex virus is attenuated. In some embodiments, the herpes simplex virus is replication-competent. In some embodiments, the herpes simplex virus is replication-deficient. In some embodiments, the herpes simplex virus is herpes simplex type 1 virus (HSV-1), herpes simplex type 2 virus (HSV-2), or any derivative thereof. In some embodiments, the herpes simplex virus is herpes simplex type 1 virus (HSV-1). In some embodiments, HSV-1 is attenuated. In some embodiments, HSV-1 has reduced cytotoxicity compared to corresponding wild-type HSV-1. In some embodiments, HSV-1 is not oncolytic.

[0325] In some embodiments, relative to the tissue tropism of unmodified wild-type herpes simplex virus, herpes simplex virus has been modified to change its tissue tropism. In some embodiments, herpes simplex virus comprises a modified envelope. In some embodiments, the modified envelope comprises one or more (e.g., one or more, two or more, three or more, four or more, etc.) mutant herpes simplex virus glycoproteins. Examples of herpes simplex virus glycoproteins may include, but are not limited to, glycoproteins gB, gC, gD, gH and gL. In some embodiments, relative to wild-type herpes simplex virus, the modified envelope changes the herpes simplex virus tissue tropism.

[0326] In some embodiments, the transduction efficiency (in vitro and / or in vivo) of a virus (e.g., herpes virus) of the present disclosure to one or more target cells (e.g., one or more human keratinocytes and / or fibroblasts) is at least about 25%. For example, the transduction efficiency of the virus to one or more target cells may be at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5% or more. In some embodiments, the virus is a herpes simplex virus, and the transduction efficiency of the virus to one or more target cells (e.g., one or more human keratinocytes and / or fibroblasts) is about 85% to about 100%. In some embodiments, the virus is a herpes simplex virus, and the transduction efficiency of the virus to one or more target cells (e.g., one or more human keratinocytes and / or fibroblasts) is at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%. Methods for measuring viral transduction efficiency in vitro or in vivo are well known to those of ordinary skill in the art, including, for example, qPCR analysis, deep sequencing, Western blot, fluorescence analysis (such as fluorescence in situ hybridization (FISH), fluorescent reporter gene expression, immunofluorescence, FACS), etc.

[0327] V. Compositions and Formulations

[0328] Certain aspects of the present disclosure relate to compositions and formulations (e.g., pharmaceutical compositions and formulations) comprising any of a recombinant nucleic acid (e.g., a recombinant herpes virus genome) and / or a virus (e.g., a herpes virus comprising a recombinant genome described herein, such as a herpes simplex virus comprising a recombinant herpes simplex virus genome) and an excipient or carrier (e.g., a pharmaceutically acceptable excipient or carrier). In some embodiments, the composition or formulation is a cosmetic composition or formulation (e.g., a skin care product).

[0329] In some embodiments, the composition or formulation comprises any one or more of the viruses described herein (e.g., herpes virus). In some embodiments, the composition or formulation comprises about 10 4 to about 10 12 Plaque forming units (PFU) / mL of virus. For example, a composition or formulation may include about 10 4 to about 10 12 , about 10 5 to about 1012 , about 10 6 to about 10 12 , about 10 7 to about 10 12 , about 10 8 to about 10 12 , about 10 9 to about 10 12 , about 10 10 to about 10 12 , about 10 11 to about 10 12 , about 10 4 to about 10 1111 , about 10 5 to about 10 11 , about 10 6 to about 10 11 , about 10 7 to about 10 11 , about 10 8 to about 10 11 , about 10 9 to about 10 11 , about 10 10 to about 10 11 , about 10 4 to about 10 10 , about 10 5 to about 10 10 , about 10 6 to about 10 10 , about 10 7 to about 10 10 , about 10 8 to about 10 10 , about 10 9 to about 10 10 , about 10 4 to about 10 9 , about 10 5 to about 10 9 , about 10 6 to about 10 9 , about 10 7 to about 10 9 , about 10 8 to about 10 9 , about 10 4 to about 10 8 , about 10 5 to about 10 8 , about 10 6 to about 108, about 10 7 to about 10 8 , about 10 4 to about 10 7 , about 10 5to about 10 7 , about 10 6 to about 10 7 , about 10 4 to about 10 6 , about 10 5 to about 10 6 , or about 10 4 to about 10 5 PFU / mL of virus. In some embodiments, the composition or formulation comprises about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 11 or about 10 12 PFU / mL of virus.

[0330] Compositions and formulations (e.g., pharmaceutical compositions and formulations) as described herein can be prepared by mixing one or more active ingredients (such as recombinant nucleic acids or viruses) having the desired purity with one or more acceptable carriers or excipients. Acceptable carriers or excipients (e.g., pharmaceutically acceptable carriers or excipients) are generally nontoxic to recipients at the dosages and concentrations used, and may include, but are not limited to: buffers (such as phosphates, citrates, acetates, and other organic acids); antioxidants (such as ascorbic acid and methionine); preservatives (such as octadecyldimethylbenzyl ammonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); amino acids (such as glycine, glutamine, asparagine, histidine, arginine, or lysine); low molecular weight (small The carrier may be a polypeptide having a length of about 10 residues; a protein such as serum albumin, gelatin, or immunoglobulin; a polyol such as glycerol, for example, a preparation comprising 10% glycerol; a hydrophilic polymer such as polyvinylpyrrolidone; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; a chelating agent such as EDTA; a sugar such as sucrose, mannitol, trehalose, or sorbitol; a salt-forming counterion such as sodium; a metal complex such as a Zn-protein complex; a liposome such as a cationic lipid; a nanoparticle carrier; and / or a nonionic surfactant such as polyethylene glycol (PEG). A thorough discussion of carriers is available in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., NJ 1991).

[0331] In some embodiments, the composition or formulation comprises one or more lipid (e.g., cationic lipid) carriers. In some embodiments, the composition or formulation comprises one or more nanoparticle carriers. Nanoparticles are submicron (less than about 1000nm) drug delivery carriers that can carry encapsulated drugs to achieve rapid or controlled release, such as synthetic small molecules, proteins, peptides, cells, viruses, and nucleic acid-based biotherapeutics. Various molecules (e.g., proteins, peptides, recombinant nucleic acids, etc.) can be efficiently encapsulated in nanoparticles using methods well known in the art. In some embodiments, molecules "encapsulated" in nanoparticles may refer to molecules (such as viruses) contained in the nanoparticles or connected to the surface of the nanoparticles and / or associated with the surface of the nanoparticles or in the form of any combination thereof. Nanoparticles used in the compositions or formulations described herein can be any type of biocompatible nanoparticles known in the art, including, for example, nanoparticles comprising poly(lactic acid), poly(glycolic acid), PLGA, PLA, PGA, and any combination thereof (see, e.g., Vauthier et al. Adv Drug Del Rev. (2003) 55:519-48; US2007 / 0148074; US2007 / 0092575; US2006 / 0246139; US5753234; US7081483; and WO2006 / 052285).

[0332] In some embodiments, the carrier or excipient (e.g., a pharmaceutically acceptable carrier or excipient) may be suitable for or adapted to any route of administration known in the art, including, for example, intravenous, intramuscular, subcutaneous, transdermal, intranasal, intratracheal, sublingual, buccal, topical, oral, transdermal, intradermal, intraperitoneal, intraorbital, intravitreal, subretinal, transmucosal, intraarticular, by superficial injection, by implantation, by inhalation, intrathecal, intraventricular, and / or intranasal administration. In some embodiments, the carrier or excipient (e.g., a pharmaceutically acceptable carrier or excipient) is suitable for or adapted to topical, transdermal, subcutaneous, and / or intradermal administration. In some embodiments, the carrier or excipient is suitable for or adapted to topical, transdermal, and / or intradermal administration. In some embodiments, the carrier or excipient is suitable for or adapted to superficial injection.

[0333] Examples of carriers or excipients suitable for or adapted for use in topical, transdermal, subcutaneous, superficial and / or intradermal application / application may include, but are not limited to, ointments, oils, pastes, creams, aerosols, suspensions, emulsions, fatty ointments, gels, powders, liquids, lotions, solutions, sprays, patches (e.g., transdermal patches or microneedle patches), adhesive tapes, microneedles or microneedle arrays, and inhalations. In some embodiments, carriers or excipients (e.g., pharmaceutically acceptable carriers or excipients) include ointments, oils, pastes, creams, aerosols, suspensions, emulsions, fatty ointments, gels, powders, liquids, lotions, solutions, sprays, adhesive tapes, and inhalations One or more (e.g., one or more, two or more, three or more, four or more, five or more, etc.). In some embodiments, carriers include patches (e.g., patches adhered to the skin), such as transdermal patches or microneedle patches. In some embodiments, carriers include microneedles or microneedle arrays. Methods for preparing and using microneedle arrays suitable for composition delivery are generally known in the art (Kim Y. et al. "Microneedles for drug and vaccine delivery". Advanced Drug Delivery Reviews 2012, 64(14): 1547-68).

[0334] In some embodiments, the composition or preparation (e.g., pharmaceutical composition or preparation) is suitable for or is suitable for any route of administration known in the art, including, for example, intravenous, intramuscular, subcutaneous, transdermal, oral, intranasal, intratracheal, sublingual, buccal, topical, transdermal, intradermal, intraperitoneal, intraorbital, intravitreal, subretinal, transmucosal, intraarticular, by superficial injection, by implantation, by inhalation, intrathecal, intraventricular and / or intranasal administration. In some embodiments, the composition or preparation is suitable for or is suitable for transdermal, topical, transdermal, subcutaneous and / or intradermal administration. In some embodiments, the pharmaceutical composition or preparation is suitable for or is suitable for topical, transdermal and / or intradermal administration. In some embodiments, the composition or preparation is suitable for or is suitable for intradermal administration. In some embodiments, the composition or preparation is suitable for or is suitable for superficial injection.

[0335] In some embodiments, a composition or formulation (eg, a pharmaceutical composition or formulation) further comprises one or more additional components. Examples of additional components may include, but are not limited to, binders (e.g., pregelatinized corn starch, polyvinyl pyrrolidone, or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates, or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silicon dioxide, colloidal silicon dioxide, stearic acid, metal stearates, hydrogenated vegetable oils, corn starch, polyethylene glycol, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium glycolate, etc.); wetting agents (e.g., sodium lauryl sulfate, etc.); salt solutions; alcohols; polyethylene glycols; gelatin; lactose; amylases; magnesium stearate; talc; silicic acid; viscous paraffin dihydroxymethylcellulose; polyvinyl pyrrolidone; sweeteners; flavorings; fragrances; colorants; humectants; opacifiers; antibacterial agents; agents capable of stabilizing polynucleotides or preventing their degradation, etc. In some embodiments, the composition or formulation comprises hydroxypropyl methylcellulose gel. In some embodiments, the composition or formulation comprises a phosphate buffer. In some embodiments, the composition or formulation comprises glycerol (e.g., at about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, etc.).

[0336] Compositions and formulations to be used for in vivo administration (eg, pharmaceutical compositions and formulations) are generally sterile. Sterility can be readily achieved, for example, by filtration through sterile filtration membranes.

[0337] In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or formulations described herein can be used to deliver one or more polynucleotides encoding collagen (e.g., human collagen such as collagen 3) to one or more cells (e.g., one or more collagen-deficient cells) of a subject. In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or formulations described herein can be used in therapy. In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or formulations described herein can be used to treat beauty or cosmetic conditions that would benefit from the expression of collagen polypeptides (e.g., beauty or cosmetic conditions associated with collagen deficiency (such as aging and / or UV-damaged skin)). In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or formulations described herein can be used to treat dermatological aging (e.g., as described below).

[0338] In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or preparations described herein can be used to prepare or manufacture a medicament. In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or preparations described herein can be used to prepare or manufacture a medicament that can be used to deliver one or more polynucleotides encoding collagen (e.g., human collagen such as collagen 3) to one or more cells (e.g., one or more collagen-deficient cells) of a subject. In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or preparations described herein can be used to prepare or manufacture a medicament that can be used to treat beauty or cosmetic conditions that will benefit from the expression of collagen polypeptides (e.g., beauty or cosmetic conditions associated with collagen deficiency (such as aging and / or UV-damaged skin)). In some embodiments, any of the recombinant nucleic acids, viruses, and / or compositions or preparations described herein can be used to prepare or manufacture a medicament that can be used to treat dermatological aging (e.g., as described below).

[0339] VI. Methods

[0340] Certain aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the level of one or more dermal extracellular matrix proteins in a subject (e.g., in one or more cells of a subject), the method comprising administering to the subject any of the recombinant nucleic acids, viruses, agents and / or compositions described herein. In some embodiments, the subject is a human.

[0341] Other aspects of the disclosure relate to methods of stabilizing or improving the structure and / or organization of a dermal extracellular matrix in a subject, the method comprising administering to the subject any of the recombinant nucleic acids, viruses, agents, and / or compositions described herein. In some embodiments, the subject is a human.

[0342] Other aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the level of one or more human collagens in a subject (e.g., in one or more cells of a subject), the method comprising administering to the subject any of the recombinant nucleic acids, viruses, agents and / or compositions described herein. In some embodiments, the subject is a human.

[0343] In some embodiments, administration of a recombinant nucleic acid, virus, agent, and / or composition to a subject increases the level (transcript or protein level) of a collagen (e.g., COL1-1; COL1-2; COL3; COL1-1 and COL1-2; COL1-1 and COL3; etc.) in one or more cells of the subject by at least about 10% compared to the endogenous level of one or more collagens in one or more corresponding untreated cells of the subject (e.g., one or more cells prior to treatment, one or more uninfected cells during treatment, etc.). For example, administration of a recombinant nucleic acid, virus, agent, and / or composition can increase the level of collagen (transcript or protein level) in one or more cells of a subject by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 99%, or more, compared to the endogenous level of one or more collagen in one or more corresponding untreated cells of the subject. In some embodiments, administration of a recombinant nucleic acid, virus, agent, and / or composition to a subject increases the level of collagen (transcript or protein level) in one or more cells of a subject by at least about 2-fold compared to the endogenous level of one or more collagen in one or more corresponding untreated cells of the subject (e.g., one or more cells before treatment, one or more uninfected cells during treatment, etc.). For example, administration of a recombinant nucleic acid, virus, agent, and / or composition can increase the level of collagen (transcript or protein level) in one or more cells of a subject by at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 50-fold, at least about 75-fold, at least about 100-fold, at least about 250-fold, at least about 500-fold, at least about 750-fold, at least about 1000-fold, or more, compared to the endogenous level of one or more collagens in one or more corresponding untreated cells of the subject. Methods for measuring transcript or protein levels from a sample are well known to those of ordinary skill in the art, including, for example, by qPCR, RNA sequencing, ELISA, Western blot, mass spectrometry, and the like.

[0344] Other aspects of the present disclosure relate to a method of enhancing, increasing, strengthening and / or supplementing the soft tissue of a subject, the method comprising administering any one of the recombinant nucleic acids, viruses, medicaments and / or compositions described herein to the subject. In some embodiments, the subject is a human. In some embodiments, the recombinant nucleic acids, viruses, medicaments and / or compositions are injected into the soft tissue of the subject. In some embodiments, the skin of the subject is aging skin. In some embodiments, the skin of the subject has been damaged due to exposure to ultraviolet light (e.g., from the sun, from tanning beds, etc.). In some embodiments, the skin of the subject is wrinkled.

[0345] In some embodiments, recombinant nucleic acids, viruses, medicaments and / or compositions can be used in methods for repairing and / or strengthening soft tissues of subjects. In some embodiments, "tissue repair" refers to restoring tissue structure and / or function, and encompasses tissue regeneration and replacement. In some embodiments, repairing or strengthening soft tissue refers to a procedure for restoring the youthful appearance of skin (e.g., "aged" skin compared to defects due to chronological aging or other physical, chemical or UV damage). In some embodiments, recombinant nucleic acids, viruses, medicaments and / or compositions can be used in cosmetic soft tissue applications, such as to fill wrinkles, lines, folds, scars, and to enhance dermal tissue (e.g., to plump up thin lips, fill sunken eyes and / or shallow cheeks, etc.).

[0346] Other aspects of the present disclosure relate to a method for improving the skin quality, condition and / or appearance of a subject in need, comprising administering to the subject any of the recombinant nucleic acids, viruses, medicaments and / or compositions described herein. In some embodiments, the subject is a human. In some embodiments, the skin condition is one or more of sun damage, aging, UV exposure, rough texture, skin sag and / or wrinkles. The improvement in skin quality, condition and / or appearance (e.g., compared to before treatment) can be assessed using any suitable method or scale known in the art including, for example, FACE Q, GAIS, etc. In some embodiments, the subject's skin is aging skin. In some embodiments, the subject's skin has been damaged due to exposure to ultraviolet light (e.g., from the sun, from tanning beds, etc.). In some embodiments, the subject's skin is wrinkled.

[0347] Other aspects of the present disclosure relate to a method for reducing the appearance of one or more superficial depressions in the skin of a subject in need, the method comprising administering any one of the recombinant nucleic acids, viruses, agents and / or compositions described herein to the subject. In some embodiments, the subject is a human. In some embodiments, the administration of recombinant nucleic acids, viruses, agents and / or compositions continues for at least about three months, at least about six months, at least about nine months or at least about 12 months to reduce the appearance of one or more superficial depressions in the skin of the subject. In some embodiments, compared to the appearance of one or more superficial depressions in the skin of the subject before the application of the composition, after the application of the composition, the appearance of one or more superficial depressions in the skin of the subject is reduced. In some embodiments, one or more superficial depressions in the skin are one or more of fine lines and wrinkles (e.g., forehead wrinkles, "crow's feet", wrinkles at the edge of the eye or mouth, etc.). In some embodiments, the treatment of one or more superficial skin depressions is measured by improvements in skin texture or skin quality such as smoothness, hydration and elasticity compared to non-treated skin. In some embodiments, treatment of one or more superficial skin depressions is measured by a reduction in the severity (e.g., depth) of the superficial depressions and / or a reduction in the number of fine lines or wrinkles in a given area of ​​the skin. In some embodiments, the subject's skin is aging skin. In some embodiments, the subject's skin has been d...

Claims

1. A use of a cosmetic composition for preparing a medicament for enhancing, increasing, strengthening and / or supplementing the level of one or more dermal extracellular matrix proteins in a subject, wherein the cosmetic composition comprises a herpes virus, wherein the herpes virus comprises a recombinant herpes virus genome, wherein the recombinant herpes virus genome comprises a first polynucleotide encoding a first cosmetic protein, and The first beauty protein is selected from the group consisting of a first collagen, a first fibronectin, a first elastin, a first aureoglycan protein, a first vitronectin, a first vitronectin receptor protein, a first laminin, a first neuromodulator protein and a first fibrillin.

2. A use of a cosmetic composition for preparing a medicament for improving the skin condition, quality and / or appearance of a subject in need thereof, wherein the cosmetic composition comprises a herpes virus, wherein the herpes virus comprises a recombinant herpes virus genome, wherein the recombinant herpes virus genome comprises a first polynucleotide encoding a first cosmetic protein, and The first beauty protein is selected from the group consisting of a first collagen, a first fibronectin, a first elastin, a first aureoglycan protein, a first vitronectin, a first vitronectin receptor protein, a first laminin, a first neuromodulator protein and a first fibrillin.

3. The use of claim 2, wherein the cosmetic composition is applied to one or more sun damaged or other UV exposed areas, rough texture areas, skin depression areas, wrinkle areas, or any combination thereof.

4. Use of a cosmetic composition for preparing a medicament for reducing the appearance of one or more superficial depressions in the skin of a subject in need thereof, the cosmetic composition comprising a herpes virus, the herpes virus comprising a recombinant herpes virus genome, wherein the recombinant herpes virus genome comprises a first polynucleotide encoding a first cosmetic protein, and The first beauty protein is selected from the group consisting of a first collagen, a first fibronectin, a first elastin, a first aureoglycan protein, a first vitronectin, a first vitronectin receptor protein, a first laminin, a first neuromodulator protein and a first fibrillin.

5. The use of claim 4, wherein the one or more superficial depressions in the skin are selected from the group consisting of nasolabial folds, crow's feet, frown lines, worry lines, scars, glabellar lines, brow ptosis, tear troughs, nasal and cheek lines, bunny lines, cheek / mid-face ptosis, marionette lines, poppy's dimples, smile lines, laugh lines, chin folds, neck lines, platysma bands, and any combination thereof.

6. Use of a cosmetic composition for preparing a medicament for increasing and / or improving at least one of the texture, smoothness, elasticity or tension of the skin of a subject in need thereof, the cosmetic composition comprising a herpes virus, the herpes virus comprising a recombinant herpes virus genome, wherein the recombinant herpes virus genome comprises a first polynucleotide encoding a first cosmetic protein, and The first beauty protein is selected from the group consisting of a first collagen, a first fibronectin, a first elastin, a first aureoglycan protein, a first vitronectin, a first vitronectin receptor protein, a first laminin, a first neuromodulator protein and a first fibrillin.

7. The use according to any one of claims 2, 4 or 6, wherein the skin of the subject is aging skin.

8. The use of any one of claims 2, 4 or 6, wherein the subject's skin has been damaged by exposure to ultraviolet light.

9. The use of any one of claims 2, 4 or 6, wherein the subject has wrinkled skin.

10. Use of a cosmetic composition for preparing a medicament for attenuating one or more dermatological signs of aging in a subject in need thereof, the cosmetic composition comprising a herpes virus, the herpes virus comprising a recombinant herpes virus genome, wherein the recombinant herpes virus genome comprises a first polynucleotide encoding a first cosmetic protein, and The first beauty protein is selected from the group consisting of a first collagen, a first fibronectin, a first elastin, a first aureoglycan protein, a first vitronectin, a first vitronectin receptor protein, a first laminin, a first neuromodulator protein and a first fibrillin.

11. The use of claim 10, wherein said reduction in one or more signs of dermatological aging is indicated by: (a) treatment, reduction and / or prevention of fine lines and / or wrinkles; (b) reduction in skin pore size; (c) improved skin thickness, fullness and / or firmness; (d) improved skin smoothness, flexibility and / or softness; (e) improved skin tone, radiance and / or clarity; (f) improved procollagen and / or collagen production; (g) improved skin texture and / or promoted retexturing; (h) improved appearance of skin contours; (i) Skin radiance and / or brightness is restored; (j) improving the appearance of skin that has diminished due to aging and / or menopause; (k) improving skin moisturization; (l) increasing skin elasticity and / or resilience; (m) treating, reducing and / or preventing skin sagging; (n) improving skin firmness; (o) reducing pigment spots, mottled skin and / or scars; (p) the optical properties of the skin with respect to light diffraction or reflection are improved; or (q) any combination thereof.

12. The cosmetic composition of any one of claims 1, 2, 4, 6 or 10, wherein the recombinant herpes virus genome is a herpes simplex virus genome.

13. The use according to claim 12, wherein the herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome.

14. The use of claim 12, wherein the recombinant herpes simplex virus gene comprises an inactivating mutation in a herpes simplex virus gene selected from the group consisting of infected cell protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41 and UL55.

15. The use according to claim 14, wherein the recombinant herpes simplex virus genome comprises an inactivating mutation in one or both copies of the ICP4 gene.

16. The use of any one of claims 1, 2, 4, 6 or 10, wherein the first cosmetic protein comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 15-21 and 53-64.

17. The use according to any one of claims 1, 2, 4, 6 or 10, wherein the first collagen is selected from the group consisting of: collagen α-1 (I) chain polypeptide (COL1-1), collagen α-2 (I) chain polypeptide (COL1-2), collagen α-1 (II) chain polypeptide (COL2), collagen α-1 (III) chain polypeptide (COL3), collagen α-1 (IV) chain polypeptide (COL4-1), collagen α-2 (IV) chain polypeptide (COL4-2), collagen α-3 (IV) chain polypeptide (COL4-3), collagen α-4 (IV) chain polypeptide (COL4-4), collagen α-5 (IV) chain polypeptide (COL4-5), collagen α-6 (I V) chain polypeptide (COL4-6), collagen alpha-1 (V) chain polypeptide (COL5-1), collagen alpha-2 (V) chain polypeptide (COL5-2), collagen alpha-3 (V) chain polypeptide (COL5-3), collagen alpha-1 (VI) chain polypeptide (COL6-1), collagen alpha-2 (VI) chain polypeptide (COL6-2), collagen alpha-3 (VI) chain polypeptide (COL6-3), collagen alpha-4 (VI) chain polypeptide (COL6-4), collagen alpha-5 (VI) chain polypeptide (COL6-5), collagen alpha-6 (VI) chain polypeptide (COL6-6), collagen alpha-1 (VIII) chain polypeptide (COL8), collagen alpha-1 (IX) chain polypeptide (COL9- 1), collagen α-2 (IX) chain polypeptide (COL9-2), collagen α-3 (IX) chain polypeptide (COL9-3), collagen α-1 (X) chain polypeptide (COL10), collagen α-1 (XI) chain polypeptide (COL11-1), collagen α-2 (XI) chain polypeptide (COL11-2), collagen α-1 (XII) chain polypeptide (COL12), collagen α-1 (XIII) chain polypeptide (COL13), collagen α-1 (XIV) chain polypeptide (COL14), collagen α-1 (XV) chain polypeptide (COL15), collagen α-1 (XVI) chain polypeptide (COL16), collagen α-1 (XVII) chain polypeptide (COL17), collagen α -1(XVIII) chain polypeptide (COL18), collagen alpha-1(XIX) chain polypeptide (COL19), collagen alpha-1(XX) chain polypeptide (COL20), collagen alpha-1(XXI) chain polypeptide (COL21), collagen alpha-1(XXII) chain polypeptide (COL22), collagen alpha-1(XXIII) chain polypeptide (COL23), collagen alpha-1(XXIV) chain polypeptide (COL24), collagen alpha-1(XXV) chain polypeptide (COL25), collagen alpha-1(XXVI) chain polypeptide (COL26), collagen alpha-1(XXVII) chain polypeptide (COL27) and collagen alpha-1(XXVIII) chain polypeptide (COL28).

18. The use according to any one of claims 1, 2, 4, 6 or 10, wherein the first collagen is selected from the group consisting of COL1-1, COL1-2, COL3, COL4-1, COL4-2, COL6-1 and COL17.

19. The use of any one of claims 1, 2, 4, 6 or 10, wherein the first collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the amino acid sequence of SEQ IDNO:

17.

20. The use of any one of claims 1, 2, 4, 6 or 10, wherein the first collagen comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO:

15.

21. The method of any one of claims 1, 2, 4, 6 or 10, wherein the recombinant herpesvirus genome does not comprise a polynucleotide encoding a collagen alpha-1 (VII) chain polypeptide (COL7), a lysyl hydroxylase 3 polypeptide (LH3) or a keratin type I cytoskeleton 17 polypeptide (KRT17).

22. The use according to any one of claims 1, 2, 4, 6 or 10, wherein the recombinant herpes virus genome further comprises a second polynucleotide encoding a second cosmetic protein.

23. The use of claim 22, wherein the first cosmetic protein and the second cosmetic protein are different.

24. The use of claim 22, wherein the first cosmetic protein and the second cosmetic protein are the same.

25. The use of any one of claims 1, 2, 4, 6 or 10, wherein the herpesvirus is replication-defective.

26. The use of any one of claims 1, 2, 4, 6 or 10, wherein the herpes virus is herpes simplex virus.

27. The use according to any one of claims 1, 2, 4, 6 or 10, wherein the cosmetic composition further comprises an excipient.

28. The use of any one of claims 1, 2, 4, 6 or 10, wherein the composition is suitable for topical, transdermal, subcutaneous, intradermal, oral, intranasal, intratracheal, sublingual, buccal, rectal, vaginal, transurethral, ​​inhalation, intravenous, intraarterial, intramuscular, intracardiac, intraosseous, intraperitoneal, transmucosal, intravitreal, subretinal, intraarticular, peri-articular, local or epidermal administration.

29. The use of any one of claims 1, 2, 4, 6 or 10, wherein the composition is suitable for intradermal administration.

30. The use of any one of claims 1, 2, 4, 6 or 10, wherein the composition is suitable for superficial injection.

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