A system for delivering genetic material subcutaneously with elements enabling subsequent post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration

Abstract

Provided herein is a system for delivering genetical material, preferably subcutaneously, and its various elements. Also provided are related compositions and methods.

Description

[0001]A SYSTEM FOR DELIVERING GENETIC MATERIAL SUBCUTANEOUSLY WITH ELEMENTS ENABLING SUBSEQUENT POST-TREATMENT IDENTIFICATION, LOCALIZATION, QUANTIFICATION, EVALUATION, OR TARGETING FOR UP- AND / OR DOWN- TITRATION RELATED APPLICATIONS This application claims the benefit of priority under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63 / 663,356, filed on June 24, 2024, the entire contents of which are incorporated herein by reference. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (R087270007WO00-SEQ- JAV.xml; Size: 29,058 bytes; and Date of Creation: June 24, 2025) is herein incorporated by reference in its entirety. FIELD OF THE INVENTION The invention relates at least in part to a system for delivering genetic material, preferably subcutaneously, and its various elements. The invention also relates at least in part to related compositions and methods. SUMMARY OF THE INVENTION In one aspect is a system, which is a gene therapy treatment for subcutaneous administration, which contains elements that enable a physical, chemical, or biological viral or non-viral delivery vector, which in an embodiment is comprised of a) one or more elements that enable the system to be redosable following initial administration and enables predictable up- titration of gene expression, b) one or more elements that enable a means of down-titration, to predictably reduce or optionally shut off the transgene expression, and c) is co-formulated with elements or co-delivered with elements, at the time of treatment, that can serve as localization markers that enable post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration. In another aspect is a physical element of delivering therapeutic transgenes, which includes one or more of electrical force, magnetic force, metallic or non-metallic particles accelerated traveling at a cell membrane penetrating velocity, physical force, or ultrasound. In an embodiment of any one of the compositions or methods provided herein, the chemical element of delivering transgenes includes an organic, inorganic, or metalloorganic molecule which can act as a surfactant, cell penetrating agent, amphiphile, or a positively charged moiety to enable transport of genetic cargo to cells and to cellular nuclei. In an embodiment of any one of the compositions or methods provided herein, the viral vector for delivering transgenes is a DNA, RNA, ssDNA, ssRNA, dsDNA, dsRNA, DNA-RNA hybrids, or a viral vector carrying a synthetic cargo. In an embodiment of any one of the compositions or methods provided herein, the viral vector for delivering transgenes is an adeno- associated viral vector, adenoviral vector, lentiviral vector, retroviral vector, HSV, or a viral vector derived from a human, mammalian, or eukaryotic viral pathogen. In an embodiment of any one of the compositions or methods provided herein, the viral vector for delivering transgenes is genetically, structurally, chemically or functionally modified, or optionally encapsulated, coated, or encapsidated into higher order structures. In an embodiment of any one of the compositions or methods provided herein, the viral vector for delivering transgenes is comprised at least in part of at least one known virus and at least one other element such as a targeting moiety, coating, formulation excipient, surfactant, lipid shell, or combination thereof. In an embodiment of any one of the compositions or methods provided herein, the non-viral vector for delivering transgenes is comprised at least in part from a lipid nanoparticle formulation. In an embodiment of any one of the compositions or methods provided herein, the non-viral vector for delivering transgenes is comprised at least in part from a polymeric nanoparticle formulation. In an embodiment of any one of the compositions or methods provided herein, the non-viral vector for delivering transgenes is comprised at least in part from a lipid nanoparticle formulation with conjugated targeting moieties. In an embodiment of any one of the compositions or methods provided herein, the non-viral vector for delivering transgenes is comprised at least in part from a polymeric nanoparticle formulation with conjugated targeting moieties. In an embodiment of any one of the compositions or methods provided herein, the non-viral vector for delivering transgenes is comprised at least in part from a formulation of virus-like particles with or without conjugated targeting moieties. In an embodiment of any one of the compositions or methods provided herein, the element that enables redosability, which is a formulation that does not elicit a neutralizing antibody response. In an embodiment of any one of the compositions or methods provided herein, the element that enables redosability, which is a formulation that locally suppresses the immune response to enable the treatment without eliciting of a neutralizing antibody response. In an embodiment of any one of the compositions or methods provided herein, the element that enables redosability, which is a non- antigenic shell, capsule, or coating of the vector or genetic cargo. In an embodiment of any one of the compositions or methods provided herein, the element that enables redosability, which is an element enabling hypoimmune characteristics of the vector and / or genetic cargo. In an embodiment of any one of the compositions or methods provided herein, the element which enables down-titration, which is a genetic cassette delivered in cis, in trans, or encoded proximally to the therapeutic transgene that enables one or more of apoptosis of the carrier cell, a suppression of protein translation, a suppression of gene expression, epigenetic self-silencing, nuclease-mediated self-inactivation, or general shut- down of gene expression reducing the level of transgene expression from the therapeutic cassette. In an embodiment of any one of the compositions or methods provided herein, the element which enables down-titration, which is an element that targets a significant proportion of the genetic cargo to adipocytes, which can be eliminated by cryolipolysis, ultrasound lipolysis, liposuction, chemical induction of adipocyte apoptosis, or pharmacological induction of adipocyte apoptosis. In an embodiment of any one of the compositions or methods provided herein, the element which enables down-titration, which is a protein expressed from the delivered genetic cassette, that enables a transient or permanent reduction in the level of gene expression following stimulation by at least one internal or external stimulus. In an embodiment of any one of the compositions or methods provided herein, the therapeutic transgene encodes at least one human or mammalian protein or peptide. In an embodiment of any one of the compositions or methods provided herein, the therapeutic transgene encodes at least an antibody. In an embodiment of any one of the compositions or methods provided herein, the therapeutic transgene encodes at least a fusion protein. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding a fluorescent protein or enzyme, which can be visualized in the fluorescent spectrum with or without inducible expression. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding one or more of at least a part of Green Fluorescent Protein (GFP), enhanced GFP, enhanced GFP with a nuclear localization signal, superfolder GFP with S30R, Y39N, N105T, Y145F, I171V, and A206V variants, emergal green GFP, Venus, mNeon Green, EYFP, Ypet, Cerulean, CyPet, AmCyan, TagBFP, EBFP, dTomato, tdTomato, DsRed Express2, mRFP1, mApple, mKate, miRFP670nano, Turbo GFP, humanized recombinant GFP, destabilized GFP, Yellow Fluorescent Protein, Cyan Fluorescent Protein, Red Fluorescent Protein, mCherry, TagRFP, mTurquoise, pHluorin2, supercliptic pH luorin, mt-mKeima, or a related protein analog, homolog, or paralog, or fragment thereof. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding a pigment protein optionally under the regulation of an inducible promoter. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding one or more of at least a fragment of bilirubin, hemocyanin, hemoglobin, myoglobin, reflectin, rhodopsin, myeloperoxidase, or other colored protein or encoding a set of DNA sequences enabling the production of pigments or pigment proteins. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding a chemiluminescent reporter optionally under the regulation of an inducible promoter. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding a reporter that is coupled with an inducer, optogenetic protein, or other regulatory element that enables its activation and visualization. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding a chromogenic reporter optionally under the regulation of an inducible promoter. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding one or more of at least a part of Firefly Luciferase, Renilla Luciferase, Gaussia Luciferase, NanoLuc Luciferase, Aequorin, Cypridina Luciferase, humanized firefly luciferase, Metridia Luciferase, Oplophorus Luciferase, Phrixothrix hirtus Luciferase, Renilla Reniformis Luciferase, or a related protein analog, homolog, or paralog, or fragment thereof. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a DNA sequence encoding a bioluminescent enzyme, which can be visualized following administration of an enzyme-specific substrate with or without inducible expression. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a pigment protein or peptide produced under the regulation of an inducible promoter. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a pigment protein or peptide produced under the regulation of a constitutive promoter. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is an imaging contrast agent. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is comprised of one or more nano- or micro-capsules, microspheres, lipid containing capsules or spheres, gadolinium based agent, metallic particles, a metal rod, sphere, or other geometric shape, a polymeric rod, sphere, or other geometric shape, a composite material rods, sphere, or other geometric shape. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is comprised of one or more biocompatible dyes. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is an adipocyte specific dye or a dye that is predominantly taken up by local subcutaneous adipocytes following administration. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a lipophilic biocompatible dye such as one or more of hematoxylin, eosin, oil red O, Congo red, alcain blue, rapid mucin, cresyl violet, Bielschowsky, fast blue, picrosirius red, Verhoeff van Gieson, Gomori’s trichrome, Masson’s trichrome, rapid PTAH, Prussian blue iron, AFB Kinyuon, AFB Ziehl- Neelson, Auramine O, Differential Quick, Fungi-Flour, Grocott Methenamine, TB Fluorostain, Warthin- Starry, Gram’s Stain, Von Kossa Dye, Villaneuva Osteochrome, Reticulin, or a biocompatible derivative or combination of biocompatible derivatives thereof. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is comprised of one or more of carbon black, titanium dioxide, silver, or silver nitrates, copper phthalocyanine, logwood extract, cobalt blue, mercury sulfide, nickel, lead carbonate, or other metalloorganic, metallic, or organic agent that can be visualized directly or following application of high sensitivity photography, imaging, or enhancing electromagnetic radiation. In an embodiment of any one of the compositions or methods provided herein, the co-formulated element, which is a die, protein, or contrast agent that is administered during the withdrawal of the administering needle to provide contrast to the dermis superficial to the site of injection, such as one or more of carbon black, titanium dioxide, copper phthalocyanine, logwood extract,cobalt blue, mercury sulfide, nickel, lead carbonate, or other metalloorganic, metallic, or organic agent that can be visualized directly or following application of high sensitivity photography, imaging, or enhancing electromagnetic radiation. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is comprised of a patch, spray, label, polymer, or liquid that is applied at the time or prior to treatment and delivers a tattooing agent to mark the position of the treatment or up-titration. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is a subcutaneously administrable marker that is opaque or partially opaque to ultrasound, or optionally palpable in the subcutis. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is a subcutaneously administrable marker that is visible under ultraviolet light, or following irradiation by a specific range of electromagnetic waves. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is an intradermally administrable marker that is opaque or partially opaque to ultrasound, or optionally palpable in the subcutis. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is an intradermally administrable marker that is visible under ultraviolet light, or following irradiation by a specific range of electromagnetic waves. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is capable of passively emitting radio waves or electromagnetic radiation following external stimulation. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is capable of storing and / or processing and / or emitting information about the originally delivered gene therapy and optionally follow-up treatments such as up- and / or down-titrations. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is capable of identifying the original location of the site of injection and optionally additional information via an RFID, Bluetooth low energy, near field communication, QR code, barcode, or other passive or active means. In an embodiment of any one of the compositions or methods provided herein, the co-delivered element, which is a tattoo superficial to the administered gene therapy injection site. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a chemical substance injected subcutaneously, intravenously, or ingested orally and enables bioluminescence of the reporter protein delivered in cis with the original therapeutic treatment. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a luciferin, fluorofurimazine, coelenterazine, cypridina luciferin analog, hikarazine, aminoluciferin, d-luciferin salt, or related substance or combination thereof. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a photoemitter with or without a coupled detected to detect and quantify fluorescence or bioluminescence from reporter protein markers delivered as part of the original gene therapy treatment. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is an ultrasound system capable of detecting the co-formulated or co-administered marker. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is one or more of a CT, MRI, SPECT, PET, infra- red, ultraviolet, or optical imaging system capable of detecting the co-formulated or co-administered marker with or without quantification. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a magnetic or electromagnetic emitter capable of detecting the co-formulated or co-delivered markers. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is an RFID reader capable of interacting with one or more elements of the delivered marker. In an embodiment of any one of the compositions or methods provided herein, the element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is an electronic device capable of interacting with and optionally parsing the information encoded in the marker via scanning of a barcode or QR code, near field communication, Bluetooth low energy, wi-fi, or other electromagnetic or radiofrequency. In an embodiment of any one of the compositions or methods provided herein, the element of enclosure of the system, which is a syringe, cartridge, vial, ampoule, or other primary container, which is optionally itself enclosed in a pen injector, autoinjector, safety syringe system, or patch injector. In another aspect is any one of the systems described herein, such as in the examples. BRIEF DESCRIPTION OF THE FIGURES Figure 1 illustrates marked injection sites by dermal tattoo. It can be confirmed to not impact reporter gene visualization by low-photon count imaging. Can use superficial camera view following substrate administration. Outputs can be signal detectable by a photon detector or camera. Down-titration can be applied to reduce level of gene expression. Down-titration can be performed by physical or pharmacological techniques. Figure 2 illustrates the effects of marking flux pre and post down- titration. Figure 3 illustrates markings added to the site of injection prior to treatment and subsequent quantification of gene expression. DETAILED DESCRIPTION OF THE INVENTION Before describing the present invention in detail, it is to be understood that this invention is not limited to particularly exemplified materials or process parameters as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only, and is not intended to be limiting of the use of alternative terminology to describe the present invention. All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety for all purposes. Such incorporation by reference is not intended to be an admission that any of the incorporated publications, patents and patent applications cited herein constitute prior art. As used in this specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the content clearly dictates otherwise. For example, reference to "a molecule" includes a mixture of two or more such molecules or a plurality of such molecules, and the like. As used herein, the term “comprise” or variations thereof such as “comprises” or “comprising” are to be read to indicate the inclusion of any recited integer (e.g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g. features, elements, characteristics, properties, method / process steps or limitations) but not the exclusion of any other integer or group of integers. Thus, as used herein, the term “comprising” is inclusive and does not exclude additional, unrecited integers or method / process steps. In embodiments of any of the compositions and methods provided herein, “comprising” may be replaced with “consisting essentially of” or “consisting of”. The phrase “consisting essentially of” is used herein to require the specified integer(s) or steps as well as those which do not materially affect the character or function of the claimed invention. As used herein, the term “consisting” is used to indicate the presence of the recited integer (e.g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g. features, elements, characteristics, properties, method / process steps or limitations) alone. Subcutaneous delivery of therapeutic transgenes enables simple accessibility from an initial route of administration perspective. The subcutis is also the main route of administration for an overwhelming majority of current recombinant protein therapies. As a tissue, the subcutis is sufficiently well vascularized to enable biodistribution of proteins delivered to, produced, and secreted in that locale, enabling systemic therapy from local protein production. Finally, adipocytes, the most prevalent resident cell of the subcutis are locally a non-life- / non-function-sustaining cell type and can enable the reduction in the dose of the gene therapy via various methods including cryolipolysis, suicide gene activation, or simple liposuction. However, the subcutaneously delivered gene therapy treatment is not visible from the surface of the skin, and generally even invasive imaging methods will not be able to pinpoint the locale of the initially delivered gene therapy. The current invention discloses elements of a gene therapy system that when delivered in tandem or co-formulated with said system can enable identification, localization, quantification, evaluation, or targeting for up- and / or down-titration of the gene therapy. Gene therapy has traditionally been delivered directly to the tissue that is missing a functional gene or via intravenous administration. As the safety profile of gene therapy treatments has improved, several attempts have been made to turn the liver or muscle into protein producing factories to augment the expression of missing factors that are normally present in circulation. While some attempts have been relatively successful the challenge with these approaches includes the turnover of the tissue in the liver, potentially requiring integrating constructs, as well as the life / function- sustaining nature of resident cells of the liver and muscle (mainly hepatocytes and myocytes), which are targeted by the gene replacement therapies. Subcutaneous delivery of therapeutic transgenes enables simple accessibility from an initial route of administration perspective. The subcutis is also the main route of administration for an overwhelming majority of current recombinant protein therapies. As a tissue, the subcutis is sufficiently well vascularized to enable biodistribution of proteins delivered to, produced, and secreted in that locale, enabling systemic therapy from local protein production. Finally, adipocytes, the most prevalent resident cells of the subcutis, are locally a non-life- / non-function- sustaining cell type and can enable the reduction in the dose of the gene therapy via various methods including cryolipolysis, suicide gene activation, or simple liposuction. At the same time, adipocytes are naturally a secretory cell, producing several proteins that have been suggested to have significant therapeutic potential including FGF21 and Apelin. As such, adipocytes and the subcutis appear to be a promising new tissue to be targeted by gene therapy. However, the subcutaneously delivered gene therapy treatment is not visible from the surface of the skin, and generally even invasive imaging methods will not be able to pinpoint the locale of the initially delivered gene therapy. Visualization of the originally administered dose can help to target it for down-titration via for example cryolipolysis, or via chemically mediated induction of adipocyte apoptosis. Moreover, non-invasive visualization of the original gene therapy treatment can enable the selection of an alternative location for subsequent up-titration. The current invention discloses elements of a gene therapy system that when delivered in tandem or co-formulated with said system can enable identification, localization, quantification, evaluation, or targeting for up- and / or down-titration of the gene therapy. The present invention generally discloses a system that is intended for the treatment of disease and is delivered subcutaneously. Said system is comprised of nucleic acids and most preferably DNA with or without one or more carriers or vectors, in a therapeutic formulation intended to treat human or veterinary pathology. The subcutaneously administered nature of the system allows it to be comprised at least in part of elements that enable downstream localization, positioning, identification, storage of data or information, or analysis for subsequent use or multiple uses. In some specific embodiments the system is a subcutaneously administered gene therapy that contains elements of one or more markers that enable the translation of information to other elements of the system that are used to assess, read, measure, identify, or locate said elements in the future, if and when required. Such second elements, which enable the localization, reading, understanding, identification, positioning, finding, or measuring of the system are generally also part of the overall gene therapy system, but are applied post-treatment to, for example, identify the original dosing site, identify the genetic cargo, measure the level of transgene expression, assess the biodistribution of the original gene therapy treatment, evaluate the required level of up- or down-titration, assess safety or efficacy of the treatment, plan future treatment protocols and paradigms, or simply locate the original site of injection for subsequent removal of the original gene therapy, dose adjustment of the original gene therapy, or re-dosing of the same or another gene therapy at an alternative site. In one general embodiment of the present invention, a system is comprised of a gene therapy treatment intended for subcutaneous administration and contains elements that enable delivery of the transgene using physical, chemical, or biological means, such as for example viral or non-viral delivery vectors, and said system is comprised of one or more elements that enable the system to be redosable following initial administration and enables predictable up- titration of gene expression, optionally includes one or more elements that enable a means of down-titration to predictably reduce or optionally shut off the transgene expression, is optionally co- formulated with elements or co-delivered with elements at the time of treatment that can serve as localization markers to enable post- treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration. In yet other embodiments the aforementioned optional elements are included in the system as critical system components to enable holistic functionality as a subcutaneously administrable gene therapy platform, with dose adjustability and markers that enable the localization of the system, its location, or evaluation following initial administration. The system may be used to deliver therapeutic transgenes, coding genes, non-coding genes, DNA, RNA, or other cargo that enables durable gene expression, durable adjustments in the level of gene expression, or other durable therapy. Said gene therapy or more specifically its cargo can be delivered using a variety of physical, chemical, or biological means. Some examples of physical delivery means including one or more of electrical force, magnetic force, metallic or non-metallic particles accelerated and traveling at a cell membrane-penetrating velocity, physical force, or ultrasound. Some examples of chemical delivery means include an organic, inorganic, or metalloorganic molecule which can act as a surfactant, cell penetrating agent, amphiphile, or a positively charged moiety to enable transport of genetic cargo to cells and to cellular nuclei. Chemical delivery can include liposomes, lipid nanoparticles, exosomes, virus like-particles, or solid lipid nanoparticles, which may optionally be assisted by physical means of delivery. Some examples of biological delivery include viral vectors, recombinant viral vectors, or modified viral vectors which may be one or more of DNA, RNA, double stranded DNA (dsDNA), single stranded DNA (ssDNA), double stranded RNA (dsRNA), single stranded RNA (ssRNA), DNA- RNA hybrids, or a viral vector carrying a synthetic cargo or modified externally with synthetic or biological molecules. Physical, chemical, and biological means of delivery may be optionally combined to assist transfection or transduction efficiency, as an example lipid nanoparticles can be conjugated with monoclonal antibodies to enhance specificity or transfection efficiency, viral particles can be encapsulated in non-antigenic synthetic shells, and either viral or non-viral particles may be assisted by physical forces during the process of delivery. Some specific viral vectors, which can be used to mediate delivery of the therapeutic cargo include adeno-associated viral vectors, adenoviral vectors, lentiviral vectors, retroviral vectors, herpes simplex virus (HSV), or a viral vector derived from a human, mammalian, or eukaryotic viral pathogen. Optionally pseudotyped or recombinant viruses can be used to deliver therapeutic cargo and said viral vectors may be additionally chemically or biologically modified to enhance tropism, specificity, safety, antigenicity, immunogenicity, or other physical, chemical or biological properties. In some specific embodiments of the present invention the viral vector used for delivering therapeutic cargo such as therapeutic transgenes may be genetically, structurally, chemically or functionally modified, or optionally encapsulated, coated, or encapsidated into higher order structures such as membranes, shells, vesicles, particles, or capsules. In yet other specific embodiments, the viral vector used for delivering therapeutic cargo such as therapeutic transgenes, which is comprised at least in part of at least one known virus and at least one other element such as a targeting moiety, coating, formulation excipient, surfactant, lipid shell, or combination thereof. Non-viral vectors may be used as part of the present invention to deliver therapeutic cargo such as transgenes for durable therapeutic transgene expression and enable redosing due to their potential non- antigenicity. Some specific non-viral vectors that can be used for this purpose include lipid nanoparticles, polymeric nanoparticles, solid lipid nanoparticles, liposomes, metallic particles, composite particles, extracellular vesicles, exosomes, or virus-like particles. Said non-viral vectors may be composed into higher order formulations to enhance stability, delivery, injectability or other physical or chemical properties and may be optionally conjugated with targeting moieties, molecules that enhance residence time, molecules that enhance stealthiness, molecules that reduce immunogenicity, molecules that reduce antigenicity, or molecules that otherwise modify chemical, physical, or biological properties of the particle or particle formulation. Similarly, virus-like particles, produced using synthetic, semi- synthetic, or recombinant methods can be used to deliver therapeutic cargo for durable therapy. Said virus-like particles can be optionally conjugated with targeting moieties, or molecules that otherwise modify the physical, chemical, or biological properties of the virus-like particle. An important element of the present invention is the non-antigenic shell or exterior of the delivery system, which under standard treatment conditions does not elicit a sufficient immune memory response to prevent subsequent redosing or up-titration of the therapy. Such an element may be generally a formulation that does not elicit a neutralizing antibody response, a formulation that locally suppresses the immune response to enable the treatment without eliciting of a neutralizing antibody response, a non-antigenic shell, capsule, capsid, envelope, membrane, monolayer, set of molecules, or coating of the vector or genetic cargo, or other element that enables hypoimmune characteristics of the vector and / or genetic cargo. Another important functional element of the system is the feature that enables down-titration or dose reduction, following initial administration. Such an element may be a genetic cassette delivered in cis, in trans, or encoded proximally to the therapeutic transgene that enables one or more of apoptosis of the carrier cell, a suppression of protein translation, a suppression of gene expression, epigenetic self- silencing, nuclease-mediated self-inactivation, or general shut-down of gene expression reducing the level of transgene expression from the therapeutic cassette. In another embodiment such an element is a feature that enables down-titration by first promoting selective targeting of the therapeutic or genetic cargo to subcutaneous adipocytes, or substantial residence in the adipose tissue following initial administration, that eventually enables elimination or reduction of gene expression via elimination or reduction of adipocytes by cryolipolysis, ultrasound lipolysis, liposuction, chemical induction of adipocyte apoptosis, or pharmacological induction of adipocyte apoptosis. In yet other embodiments an element that enables down-titration or dose reduction may be comprised at least in part from a responsive or inducible promoter or at least in part of a protein expressed from the delivered genetic cassette, that enables a transient or permanent reduction in the level of gene expression following stimulation by at least one internal or external stimulus. Some examples include the tetracycline-inducible promoter, or the doxycycline-inducible promoter, or a DNA-binding repressor protein, or an RNA expressed following activation that reduces the level of gene expression. Said down-titration is ideally permanent and predictable, but may be transient in nature, such that the level of expression recovers following the removal of the stimulus. The therapeutic formulation described in the present invention is intended to deliver durable therapy, which has an adjustable level of therapeutic effect. Such durable therapy may be optionally achieved using non-integrating DNA cassettes, or optionally integrating DNA cassettes or transposable or retrotransposable elements. Said elements may contain one or more therapeutic genes, which may be coding or non- coding genes that express one or more proteins or one or more RNAs. Such therapeutic genes may be comprised of one or more genes that encode at least one human or mammalian protein or peptide. In another embodiment, the protein or peptide is secreted to enable production in adipocytes and secretion into the general circulation. Some such proteins may include one or more antibodies, fusion proteins, peptides, or combinations thereof. A central element of the present invention is a co-formulated element or a marker, indicator, or locator, that is comprised of at least a part of some physical, chemical, or biological component and allows for the localization, identification, obtainment of information from, or positioning of the subcutaneously administered treatment for some subsequent action upon that treatment or related to the treatment. In some embodiments such an element may be a marker for identification of the location of the original treatment, in other embodiments the indicator may serve as a means of providing information about the treatment such as its activity, in other embodiment the co-formulated element may be only visualized or apparent following application of one or more stimuli and may be used for location, identification, quantification, collection of information from, or a combination of one or more actions upon or related to the original treatment. As an example, the marker may be used to identify the location of the treatment for subsequent down-titration, or optionally in conjunction with the location provide information about the activity of the treatment or the level of therapeutic gene expression at the site of original treatment, this information can be used to enable down- titration or up-titration, or the selection of an action which is neither down-titration or up- titration by the treating physician, the patient, or the technician performing the measurement or evaluation. One such co-formulated element delivered together with the original treatment can be comprised of at least in part of a DNA sequence encoding a fluorescent protein or enzyme, which can be visualized in the fluorescent spectrum with or without inducible expression. Another such co-formulated element can be comprised at least in part of a DNA sequence encoding one or more of at least a part of Green Fluorescent Protein (GFP), enhanced GFP, enhanced GFP with a nuclear localization signal, superfolder GFP with S30R, Y39N, N105T, Y145F, I171V, and A206V variants, emerald green GFP, Venus, mNeon Green, EYFP, Ypet, Cerulean, CyPet, AmCyan, TagBFP, EBFP, dTomato, tdTomato, DsRed Express2, mRFP1, mApple, mKate, miRFP670nano, Turbo GFP, humanized recombinant GFP, destabilized GFP, Yellow Fluorescent Protein, Cyan Fluorescent Protein, Red Fluorescent Protein, mCherry, TagRFP, mTurquoise, pHluorin2, supercliptic pH luorin, mt-mKeima, or a related protein analog, homolog, or paralog, or fragment thereof. Said proteins can be constituitively or inducibly expressed and can be used in conjunction with other tools such as high-sensitivity cameras or detectors to identify the location of treatment, measure the level of activity or transgene expression, quantify the durability of the treatment, potential efficacy, or potential need for up- or down- titration and optionally the target site of the up- or down-titration application. Another such co-formulated element can be comprised of one or more of a DNA sequence encoding a pigment protein, pigment proteins, or parts of pigment proteins optionally under the regulation of an inducible promoter. Other such co-formulated elements may be comprised of one or more DNA sequences encoding one or more of at least a fragment of bilirubin, hemocyanin, hemoglobin, myoglobin, reflectin, rhodopsin, myeloperoxidase, or other colored protein or encoding a set of DNA sequences enabling the production of pigments or pigment proteins. Yet other such co-formulated elements can be comprised of one or more DNA sequences encoding a chemiluminescent reporter optionally under the regulation of an inducible promoter. And yet other such co- formulated elements can be comprised of one or more DNA sequences encoding a reporter that is coupled with an inducer, optogenetic protein, or other regulatory element that enables its activation and visualization. In some embodiments of the present invention the co- formulated element may be a DNA sequence encoding a chromogenic reporter optionally under the regulation of an inducible promoter, or a DNA sequence encoding one or more of at least a part of Firefly Luciferase, Renilla Luciferase, Gaussia Luciferase, NanoLuc Luciferase, Aequorin, Cypridina Luciferase, humanized firefly luciferase, Metridia Luciferase, Oplophorus Luciferase, Phrixothrix hirtus Luciferase, Renilla Reniformis Luciferase, or a related protein analog, homolog, or paralog, or fragment thereof, or one or more DNA sequences encoding one or more similar proteins, peptides, or fusion proteins, or proteins encoding pathway proteins required to produce non-protein pigments such as for example melanin. In yet other embodiments of the present invention the co- formulated element cam be comprised at least in part of a DNA sequence encoding a bioluminescent enzyme, which can be visualized following administration of an enzyme-specific substrate with or without inducible expression. Said co-formulate elements may be regulated by a cell-specific promoter, an inducible promoter sequence, a circadian rhythm promoter, a dual-inducible promoter sequence, a responsive promoter such as for example a heat-shock responsive promoter, or a constitutive promoter. A co-formulated element delivered to enable localization, visualization, dose adjustment, dose quantification, or other factor associated with the treatment may be a pigment protein or peptide, or a set of proteins responsible for producing a non-protein pigment or color, light, increased density, increased opacity, or electromagnetic radiation by one or more means and may be under the regulation of one or more constitutive or inducible promoters. In some embodiments of the present invention the original therapeutic formulation contains a co-formulated element, which is a standard imaging contrast agent that can be used for visualization, localization, or identification of the original treatment location. In some specific embodiments the co-formulated element is comprised of one or more nano- or micro-capsules, microspheres, lipid containing capsules or spheres, gadolinium-based agent, metallic particles, a metal rod, sphere, or other geometric shape, a polymeric rod, sphere, or other geometric shape, a composite material rods, sphere, or other geometric shape. In some other specific embodiments a co-formulated agent administered during the original treatment is comprised at least in part of one or more biocompatible dyes. In yet other embodiments a co-formulated agent maybe administered immediately after the original injection from a formulation that is split across for example to primary containers or a primary container that is comprised of at least two chambers, or one chamber that is at least partially physically separated from another. Co-formulated agents may be dyes, pigments, contrast agents or other physical, chemical, or biological elements that are at least partially adipocyte-specific or adipocyte-targeting, and are taken up by adipocytes, or predominantly taken up by adipocytes, or predominantly taken up by tissues in the location of the original injection without substantial translocation away from the original site for at least 1 year following initial administration. In some specific embodiments of the present invention the co-formulated element is comprised at least in part from at least a part of a lipophilic biocompatible dye such as one or more of hematoxylin, eosin, oil red O, Congo red, alcain blue, rapid mucin, cresyl violet, Bielschowsky, fast blue, picrosirius red, Verhoeff van Gieson, Gomori’s trichrome, Masson’s trichrome, rapid PTAH, Prussian blue iron, AFB Kinyuon, AFB Ziehl- Neelson, Auramine O, Differential Quick, Fungi-Flour, Grocott Methenamine, TB Fluorostain, Warthin-Starry, Gram’s Stain, Von Kossa Dye, Villaneuva Osteochrome, Reticulin, or a biocompatible derivative or combination of biocompatible derivatives thereof. In some specific embodiments of the present invention the co- formulated element may be comprised at least in part of at least a part of one or more of carbon black, titanium dioxide, silver, or silver nitrates, copper phthalocyanine, logwood extract, cobalt blue, mercury sulfide, nickel, lead carbonate, or other metalloorganic, metallic, or organic agent that can be visualized directly or following application of high sensitivity photography, imaging, or enhancing electromagnetic radiation. And yet in other specific embodiments of the present invention the co-formulated element may be comprised at least in part of a die, protein, or contrast agent that is administered during the withdrawal of the administering needle to provide contrast to the dermis superficial to the site of injection, such as one or more of carbon black, titanium dioxide, copper phthalocyanine, logwood extract, cobalt blue, mercury sulfide, nickel, lead carbonate, or other metalloorganic, metallic, or organic agent that can be visualized directly or following application of high sensitivity photography, imaging, or enhancing electromagnetic radiation. Multiple co-formulated elements may be utilized together, for example a gene expressing a bioluminescent protein to help in the quantification of gene expression from the original treatment with the help of a detector and bioluminescent substrate can be used together with a pigment or pigment protein. In this case the former enables measurement of gene expression from the original treatment and the latter enables localization of the treatment for the enablement of down-titration, redosing, up-titration, shutdown of gene expression, or the selection of an alternative treatment site for another therapy. The therapy described in the present invention may be comprised of a formulation, multiple formulations, or formulation or multiple formulations together with other components that contain co-delivered elements. Said co-delivered elements may be comprised of one or more of a patch, spray, label, polymer, or liquid that is applied at the time or prior to treatment and delivers a tattooing agent to mark the position of the treatment or up-titration. In other embodiments said co-delivered elements may be comprised at least in part of a subcutaneously administrable marker that is opaque or partially opaque to ultrasound, or optionally palpable in the subcutis. And in yet other embodiments of the present invention said co-delivered elements may be comprised at least in part of a subcutaneously administrable marker that is visible under ultraviolet light, or following irradiation by a specific range of electromagnetic waves. Any co-delivered elements can be delivered before, during, or after the administration of the original therapeutic treatment and should be ideally delivered to the site of treatment, or optionally proximal to the treatment such that they are capable of providing a future indication of the original treatment location. In some specific embodiments of the present invention the co-delivered element can be comprised at least in part of an intradermally administrable marker that is opaque or partially opaque to ultrasound, or optionally palpable in the subcutis. In other specific embodiments of the present invention the co-delivered element can be comprised at least in part of an intradermally administrable marker that is visible under ultraviolet light, or following irradiation by a specific range of electromagnetic waves. Optionally a co-delivered element may be comprised of an entity which is capable of passively emitting radio waves or electromagnetic radiation following external stimulation, or one which is capable of storing and / or processing and / or emitting information about the originally delivered gene therapy and optionally follow-up treatments such as up- and / or down-titrations. Co-delivered elements may be enabled with functionality the provides a means of identifying the original location of the site of injection and optionally additional information via an RFID, Bluetooth low energy, near field communication, QR code, barcode, or other passive or active means. In yet other embodiments of the present invention the co- delivered element may be comprised at least in part of a tattoo superficial to the administered gene therapy injection site. The system delivered for therapeutic effect is comprised of one or more elements that allow for future functionality of said system such as its identification, localization, quantification, or collection of information. This functionality is enabled by elements co- administered, co-delivered, or co-formulated with the system, but the functionality may be afforded to the system by said elements at the time of treatment or after the original treatment. Other elements may be required to enable this functionality at the time of treatment or following initial treatment. Some examples of said elements that enable future functionality include for example substrates for bioluminescent reporter proteins co-expressed from the in-cis delivered genetic cassettes, or cameras required for the detection and quantification of expression of said reporter proteins or the photons produced from said reporter proteins. In some embodiments such an element used as part of said functionality of the system, but optionally delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a chemical substance injected subcutaneously, intravenously, or ingested orally and enables bioluminescence of the reporter protein delivered in cis with the original therapeutic treatment. Alternatively, such an element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, is comprised at least in part of at least a part of a luciferin, fluorofurimazine, coelenterazine, cypridina luciferin analog, aminoluciferin, d-luciferin salt, or related substance or combination thereof. Other such elements that can enable some functionality of the system include one or more of a photoemitter with or without a coupled detected to detect and quantify fluorescence or bioluminescence from reporter protein markers delivered as part of the original gene therapy treatment, ultrasound system capable of detecting the co-formulated or co-administered marker, a CT, MRI, SPECT, PET, infra-red, ultraviolet, or optical imaging system capable of detecting the co-formulated or co-administered marker with or without quantification. In yet other embodiments of the present invention an element or elements used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration can be comprised at least a part from magnetic or electromagnetic emitter capable of detecting the co-formulated or co-delivered markers. In yet other embodiments of the present invention said elements for the enablement of functionality can be comprised of an RFID reader capable of interacting with one or more elements of the delivered marker. And in yet other embodiments of the present invention said elements can be comprised at least in part of a barcode or QR code, near field communication, Bluetooth low energy, wi-fi, or other electromagnetic or radiofrequency. Said elements may be used post treatment to localize, detect, quantify, or collect other information about the original treatment to enable its localization, assessment, evaluation, identification, or generally to enable decision-making about future treatment, up-titration, redosing, or down-titration or dose reduction. As an example a camera detecting photons emitted from a bioluminescent reporter protein following administration of a substrate locally to the original injection site, can provide information about the level of gene expression at some period post initial treatment and inform a decision about potential up-titration or down-titration of treatment, for example whether to deliver an augmenting dose or up-titrate, to leave the dose unchanged as the level of reporter gene expression may be assessed as optimal or sufficient relative to some early or original measurement, or to down- titrate treatment by for example cryolipolysis if required. The system described in the present invention may be additionally comprised of a primary and secondary container, or generally an enclosure system, which is a syringe, cartridge, vial, ampoule, or other primary container, which is optionally itself enclosed in a pen injector, autoinjector, safety syringe system, or patch injector. Some specific examples of enabling the present invention are outlined in detail in the example section, but are not intended to be limiting to the scope of the invention. Elements of the invention may be varied by someone trained in the art, combined with other elements or parts of other elements to enhance functionality with the ultimate goal of embodying a gene therapy system that has one or more marking elements. Nucleic acid sequences can be codon optimized using general codon optimization tools using artificial intelligence-based tools and specifically for a given species of interest. Auxiliary sequences may be delivered in cis or in trans with the nucleic acid sequences encoding the therapeutic protein of interest and the secretion signal and may include one or more of promoters, terminators, insulators, 3’UTRs, 5’UTRs, enhancers, or combinations thereof. The nucleic acid sequences encoding the markers / reporter proteins, or the therapeutic peptide or protein of interest which are intended for the treatment of human or veterinary pathologies and may be generally derived from natural sequences, synthetic sequences, or semi-synthetic sequences with or without in silico optimization, with or without the use of codon optimization, with or without the use of artificial intelligence tools or systems and with or without the use of directed evolution. It should be understood that within the scope of this invention the amino acid sequences, nucleic acid sequences, motifs, biological and chemical compositions, or methods of use may be varied by one skilled in the art, to the extent that the structures described herewithin perform the desired function and remain within the scope of the present invention. Various parts, components or characteristics may be used in combination, with or without modification by someone skilled in the art to achieve the desired functionality of the aforedescribed formulation. Moreover, all individual features and methods of use described herein, and each and every combination of two or more of such features and methods of use, are included within the scope of the present invention provided that these features and methods of use in such a combination are not mutually inconsistent. It is understood that certain portions or combinations of such portions can be varied by someone trained in the art while still achieving the main goal of the invention. Finally, it is understood that the specific ranges provided in the current invention are not restrictive and are for example purposes only, values outside of the specified ranges may be used to achieve the goal of the invention without modification to the proposed mechanistic principals. EXAMPLES Example 1 In the present example, a gene therapy treatment for type 2 diabetes and obesity is administered using a lipid nanoparticle vector. The vector contains a DNA cassette optimized to remove TLR-reactive sequences and intracellular DNA sensors, include nuclear targeting sequences, and encodes a natural human GLP-1 sequence modified for enhanced resistance to peptidase digestion. The GLP-1 is expressed under the regulation of the CAG promoter, specifically containing (C) the cytomegalovirus (CMV) early enhancer element, (A) the promoter, the first exon and the first intron of chicken beta-actin gene, and (G) the splice acceptor of the rabbit beta-globin gene. The genetic cassette additionally encodes a low-leak tetracycline inducible promoter, which drives the expression of a humanized recombinant green fluorescent protein derived from Renilla reniformis. Following initial treatment, upon the desire to evaluate the number of genetic cassettes using bioluminescence as a surrogate indicator the quantity of expressed luciferase is measured by first inducing expression with a fixed dose of oral tetracycline, followed by the irradiation of the abdomen with a device emitting a narrow band path filtered flood light with a peak emission of 500 nm, which is coupled with a detector with narrow bandpass filter of max transmittance at 506 nm. An optional measurement is performed 1 day, 7 days, 30 days and 60 days post initial dosing and followed up with additional quantifications as required for the optimization of the safety or efficacy of the treatment. Should redosing be required, the identification of the reporter gene position can enable the selection of a second subcutaneous dose treatment site at a distance greater than 1.0 cm from the original location. Should the reduction in the therapeutic transgene expression be required, the administration of a down-titration mechanisms such as cryolipolysis can be performed to the site of the original injection as identified by the expressed marker. TRE promoter GGTACCGAGCTCGACTTTCACTTTTCTCTATCACTGATAGGGAGTGGTAAACTCGACTTTCACTTTTC TC TATCACTGATAGGGAGTGGTAAACTCGACTTTCACTTTTCTCTATCACTGATAGGGAGTGGTAAACTC GA CTTTCACTTTTCTCTATCACTGATAGGGAGTGGTAAACTCGACTTTCACTTTTCTCTATCACTGATAG GG AGTGGTAAACTCGACTTTCACTTTTCTCTATCACTGATAGGGAGTGGTAAACTCGACTTTCACTTTTC TC TATCACTGATAGGGAGTGGTAAACTCGACCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGC CT GGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCCGCGGCCCC GA ATTG (SEQ ID NO: 1) RtTA response element ATGTCTAGACTGGACAAGAGCAAAGTCATAAACGGCGCTCTGGAATTACTCAATGGAGTCGGTATCGAAG GCCTGACGACAAGGAAACTCGCTCAAAAGCTGGGAGTTGAGCAGCCTACCCTGTACTGGCACGTGAAGAA CAAGCGGGCCCTGCTCGATGCCCTGCCAATCGAGATGCTGGACAGGCATCATACCCACTTCTGCCCCCTG GAAGGCGAGTCATGGCAAGACTTTCTGCGGAACAACGCCAAGTCATTCCGCTGTGCTCTCCTCTCACATC GCGACGGGGCTAAAGTGCATCTCGGCACCCGCCCAACAGAGAAACAGTACGAAACCCTGGAAAATCAGCT CGCGTTCCTGTGTCAGCAAGGCTTCTCCCTGGAGAACGCACTGTACGCTCTGTCCGCCGTGGGCCACTTT ACACTGGGCTGCGTATTGGAGGAACAGGAGCATCAAGTAGCAAAAGAGGAAAGAGAGACACCTACCACCG ATTCTATGCCCCCACTTCTGAGACAAGCAATTGAGCTGTTCGACCGGCAGGGAGCCGAACCTGCCTTCCT TTTCGGCCTGGAACTAATCATATGTGGCCTGGAGAAACAGCTAAAGTGCGAAAGCGGCGGGCCGGCCGAC GCCCTTGACGATTTTGACTTAGACATGCTCCCAGCCGATGCCCTTGACGACTTTGACCTTGATATGCTGC CTGCTGACGCTCTTGACGATTTTGACCTTGACATGCTCCCCGGGTAA (SEQ ID NO: 2) Which is followed by an SV40 late polyA CAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTT AT TTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACA AC AATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCT CT ACAAATGTGGTA (SEQ ID NO: 3) Humanized efficient Green Fluorescent Protein (heGFP) ATGGTGAGCAAGCAGATCCTGAAGAACACCGGCCTGCAGGAGATCATGAGCTTCAAGGTGAACCTGGA GG GCGTGGTGAACAACCACGTGTTCACCATGGAGGGCTGCGGCAAGGGCAACATCCTGTTCGGCAACCAG CT GGTGCAGATCCGCGTGACCAAGGGCGCCCCCCTGCCCTTCGCCTTCGACATCCTGAGCCCCGCCTTCC AG TACGGCAACCGCACCTTCACCAAGTACCCCGAGGACATCAGCGACTTCTTCATCCAGAGCTTCCCCGC CG GCTTCGTGTACGAGCGCACCCTGCGCTACGAGGACGGCGGCCTGGTGGAGATCCGCAGCGACATCAAC CT GATCGAGGAGATGTTCGTGTACCGCGTGGAGTACAAGGGCCGCAACTTCCCCAACGACGGCCCCGTGA TG AAGAAGACCATCACCGGCCTGCAGCCCAGCTTCGAGGTGGTGTACATGAACGACGGCGTGCTGGTGGG CC AGGTGATCCTGGTGTACCGCCTGAACAGCGGCAAGTTCTACAGCTGCCACATGCGCACCCTGATGAAG AG CAAGGGCGTGGTGAAGGACTTCCCCGAGTACCACTTCATCCAGCACCGCCTGGAGAAGACCTACGTGG AG GACGGCGGCTTCGTGGAGCAGCACGAGACCGCCATCGCCCAGCTGACCAGCCTGGGCAAGCCCCTGGG CA GCCTGCACGAGTGGGTGTAA (SEQ ID NO: 4) CAG promoter CTCGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATAT AT GGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCAT TG ACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGA GT ATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGAC GT CAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGC AG TACATCTACGTATTAGTCATCGCTATTACCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCC CA TCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTTTGTGCAGCGATGGGG GC GGGGGGGGGGGGGGGGCGCGCGCCAGGCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGA GA GGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGCGGCGGCGGCG GC GGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGGGAGTCGCTGCGCGCTGCCTTCGCCCCGTGCCCCGC TC CGCCGCCGCCTCGCGCCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGG AC GGCCCTTCTCCTCCGGGCTGTAATTAGCGCTTGGTTTAATGACGGCTTGTTTCTTTTCTGTGGCTGCG TG AAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGAGCGGCTCGGGGGGTGCGTGCGTGTGT GT GTGCGTGGGGAGCGCCGCGTGCGGCTCCGCGCTGCCCGGCGGCTGTGAGCGCTGCGGGCGCGGCGCGG GG CTTTGTGCGCTCCGCAGTGTGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGG CT GCGAGGGGAACAAAGGCTGCGTGCGGGGTGTGTGCGTGGGGGGGTGAGCAGGGGGTGTGGGCGCGTCG GT CGGGCTGCAACCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCACGGCCCGGCTTCGGGTGCGGGGC TC CGTACGGGGCGTGGCGCGGGGCTCGCCGTGCCGGGCGGGGGGTGGCGGCAGGTGGGGGTGCCGGGCGG GG CGGGGCCGCCTCGGGCCGGGGAGGGCTCGGGGGAGGGGCGCGGCGGCCCCCGGAGCGCCGGCGGCTGT CG AGGCGCGGCGAGCCGCAGCCATTGCCTTTTATGGTAATCGTGCGAGAGGGCGCAGGGACTTCCTTTGT CC CAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCCTCTAGCGGGCGCGGGGCGAAGCG GT GCGGCGCCGGCAGGAAGGAAATGGGCGGGGAGGGCCTTCGTGCGTCGCCGCGCCGCCGTCCCCTTCTC CC TCTCCAGCCTCGGGGCTGTCCGCGGGGGGACGGCTGCCTTCGGGGGGGACGGGGCAGGGCGGGGTTCG GC TTCTGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACA GC TCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTTGGCAAAGAATTG (SEQ ID NO: 5) Example 2 In the present example a gene therapy treatment is administered using an AAV vector delivering an IL-1 receptor antagonist for subcutaneous expression to treat rheumatoid arthritis. At the time of treatment, a machine-readable fluorescein tattoo is applied at the injection site to encode the therapeutic target (IL-1Ra), dose, and treatment date. The center of the machine-readable image forms the center of the site of injection. At the time of required up-titration, the approximate injection area is illuminated with a 490-495 nm UV lamp to locate the original injection site and verify the treatment and dose. The information is used to identify an alternative location for a subsequent injection and assess the required dose based on the safety and efficacy profile of the original treatment. At the time of required down-titration, the approximate injection area is illuminated with a 490-495 nm UV lamp to locate the original injection site and verify the treatment and dose. The information is used to target the down-titration treatment, which is a subcutaneous injection of a lipolytic agent. Upon verification of the elimination of the required therapy via verification of therapeutic transgene expression using serological biomarkers, the machine-readable fluorescein tattoo is removed via the application of 1064 and 532 nm lasers, followed by verification of removal using a 490-495 nm UV light source. IL-1Ra sequence RPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKIDVVPIEPHALFLGIHGGKMCLS CV KSGDETRLQLEAVNITDLSENRKQDKRFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPD EG VMVTKFYFQEDE (SEQ ID NO: 6) Example 3 A polymeric nanoparticle vector is used to deliver a genetic cassette that encodes the natural human Apelin agonist for the treatment of sarcopenia and muscle wasting disorders. The treatment is administered subcutaneously and delivers the therapeutic transgene under the regulation of a constitutive promoter, as well as a near-infrared (NIR) fluorescent protein derived from the cyanobacteriochrome photoreceptor NpR3784 (miRFP670nano). Following treatment, oral tetracycline can be used to induce expression of the near-infrared fluorescent protein and an NIR camera can be used to detect the location of the original site of administration and strength of gene expression. As the reporter remains as an intracellular protein and is expressed from the same genetic cassette, the signal strength can be used as a surrogate measure of the total genome copies of the therapeutic construct present in the treatment area. Such quantification can be performed at various timepoints to assess the level of gene expression and drive further treatment decisions. The NIR signal can be additionally used to localize the original site of injection for subsequent down-titration for example by cryolipolysis. Cryolipolytic treatment can be applied overtop of the NIR signal and optionally to areas immediately adjacent to the NIR signal. Similarly, the signal location can be also used to select alternative sites for subsequent gene therapy administration, or re- dosing to up-titrate the initial dose should additional therapeutic efficacy be required for a particular patient. MiRFP670nano ATGGCAAACCTGGACAAGATGCTGAATACCACAGTAACAGAGGTGCGGCAGTTCCTGCAGGTGGACAGA G TGTGCGTGTTCCAGTTTGAGGAGGATTATAGCGGAGTGGTGGTGGTGGAGGCCGTGGACGATAGGTGGA T CTCCATCCTGAAGACCCAGGTGCGGGATAGATACTTCATGGAGACAAGGGGCGAGGAGTATTCTCACGG C CGCTACCAGGCCATCGCCGACATCTACACCGCAAACCTGACAGAGTGCTACAGGGATCTGCTGACACAG T TTCAGGTGAGAGCAATCCTGGCCGTGCCCATCCTGCAGGGCAAGAAGCTGTGGGGCCTGTTGGTGGCAC A CCAGCTGGCGGCCCCTAGACAGTGGCAGACCTGGGAGATCGACTTTCTGAAGCAGCAGGCCGTGGTGGT G GGCATCGCCATCCAGCAGAGCTAG (SEQ ID NO: 7) Apelin ATGAATCTGCGGCTCTGCGTGCAGGCGCTCCTGCTGCTCTGGCTCTCCTTGACCGCGGTGTGTGGAGG GT CCCTGATGCCGCTTCCCGATGGGAATGGGCTGGAAGACGGCAATGTCCGCCACCTGGTGCAGCCCAGA GG GTCAAGGAATGGGCCAGGGCCCTGGCAGGGAGGTCGGAGGAAATTCCGCCGCCAGCGGCCCCGCCTCT CC CATAAGGGACCCATGCCTTTCTGA (SEQ ID NO: 8) TTS ATGTCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGT TT AACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCACGTGCGCAACAAGCA GA CTCTTATGAACATGCTTTCAGAGGCAATACTGGCGAAGCATCACACCCGTTCAGCACCGTTACCGACTGAGA GT TGGCAGCAGTTTCTCCAGGAAAATGCTCTGAGTTTCCGTAAAGCATTACTGGTCCATCGTGATGGAGCCCGA TT GCATATAGGGACCTCTCCTACGCCCCCCCAGTTTGAACAAGCAGAGGCGCAACTACGCTGTCTATGCGATGC AG GGTTTTCGGTCGAGGAGGCTCTTTTCATTCTGCAATCTATCAGCCATTTTACGTTGGGTGCAGTATTAGAGG AG CAAGCAACAAACCAGATAGAAAATAATCATGTGATAGACGCTGCACCACCATTATTACAAGAGGCATTTAAT AT TCAGGCGAGAACCTCTGCTGAAATGGCCTTCCATTTCGGGCTGAAATCATTAATATTTGGATTTTCTGCACA GT TAGATGAAAAAAAGCATACACCCATTGAGGATGGTAATAAACCAAAAAAGAAGAGAAAGCTAGCAGTGTCAG TG ACATTTGAAGATGTGGCTGTGCTCTTTACTCGGGACGAGTGGAAGAAGCTGGATCTGTCTCAGAGAAGCCTG TA CCGTGAGGTGATGCTGGAGAATTACAGCAACCTGGCCTCCATGGCAGGATTCCTGTTTACCAAACCAAAGGT GA TCTCCCTGTTGCAGCAAGGAGAGGATCCCTGG (SEQ ID NO: 9) T2A GAGGGCAGGGGAAGTCTTCTAACATGCGGGGACGTGGAGGAAAATCCCGGCCCC (SEQ ID NO: 10) RtTA ATGTCTAGACTGGACAAGAGCAAAGTCATAAACGGCGCTCTGGAATTACTCAATGGAGTCGGTATCGAA G GCCTGACGACAAGGAAACTCGCTCAAAAGCTGGGAGTTGAGCAGCCTACCCTGTACTGGCACGTGAAGA A CAAGCGGGCCCTGCTCGATGCCCTGCCAATCGAGATGCTGGACAGGCATCATACCCACTTCTGCCCCCT G GAAGGCGAGTCATGGCAAGACTTTCTGCGGAACAACGCCAAGTCATTCCGCTGTGCTCTCCTCTCACAT C GCGACGGGGCTAAAGTGCATCTCGGCACCCGCCCAACAGAGAAACAGTACGAAACCCTGGAAAATCAGC T CGCGTTCCTGTGTCAGCAAGGCTTCTCCCTGGAGAACGCACTGTACGCTCTGTCCGCCGTGGGCCACTT T ACACTGGGCTGCGTATTGGAGGAACAGGAGCATCAAGTAGCAAAAGAGGAAAGAGAGACACCTACCACC G ATTCTATGCCCCCACTTCTGAGACAAGCAATTGAGCTGTTCGACCGGCAGGGAGCCGAACCTGCCTTCC T TTTCGGCCTGGAACTAATCATATGTGGCCTGGAGAAACAGCTAAAGTGCGAAAGCGGCGGGCCGGCCGA C GCCCTTGACGATTTTGACTTAGACATGCTCCCAGCCGATGCCCTTGACGACTTTGACCTTGATATGCTG C CTGCTGACGCTCTTGACGATTTTGACCTTGACATGCTCCCCGGGTAA (SEQ ID NO: 11) BHG polyA CTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCA CT CCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGG GG TGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTC TA TGG (SEQ ID NO: 12) Example 4 A gene therapy treatment is administered subcutaneously to target white adipocytes using a lipid nanoparticle formulation delivering a genetic cassette that encodes human FGF21 for the treatment of Metabolic dysfunction–associated steatotic liver disease (MASLD). The genetic cassette additionally encodes a TRE promoter, that conditionally drives expression of an inducible suicide gene (iCasp9) following administration of tetracycline. The FGF21 therapeutic transgene is driven by the UBC promoter and followed by the rBG polyA signal. At the time of treatment an RFID chip is implanted subcutaneously at the site of treatment, but sufficiently superficially to be palpable through the dermis and epidermis. At the time of treatment the RFID chip is populated with the following information: treatment type, therapeutic transgene type, administered dose in terms of total genome copies delivered, total injection volume, date of treatment, name or number of treating physician, condition undergoing treatment, and additional alphanumeric variable data of the physician’s choice. At time of required dose adjustment, or at time of therapy review, the above information is scanned into an RFID reader to enable analysis or further treatment decisions. Should up- or down-titration of the therapeutic dose be required, the RFID is used to locate the original site of injection using palpation or proximity sensing devices. Up- titration is applied at a site distal to the original injection by at least 1.5 cm and down-titration is applied via the injection to the original site of administration a combination of tetracycline and rapamycin, or a chemical lipolytic agent. Upon the administration of down-titration the RFID is optionally explanted. FGF21 sequence ATGGACTCGGACGAGACCGGGTTCGAGCACTCAGGACTGTGGGTTTCTGTGCTGGCTGGTCTTCTGCT GG GAGCCTGCCAGGCACACCCCATCCCTGACTCCAGTCCTCTCCTGCAATTCGGGGGCCAAGTCCGGCAG CG GTACCTCTACACAGATGATGCCCAGCAGACAGAAGCCCACCTGGAGATCAGGGAGGATGGGACGGTGG GG GGCGCTGCTGACCAGAGCCCCGAAAGTCTCCTGCAGCTGAAAGCCTTGAAGCCGGGAGTTATTCAAAT CT TGGGAGTCAAGACATCCAGGTTCCTGTGCCAGCGGCCAGATGGGGCCCTGTATGGATCGCTCCACTTT GA CCCTGAGGCCTGCAGCTTCCGGGAGCTGCTTCTTGAGGACGGATACAATGTTTACCAGTCCGAAGCCC AC GGCCTCCCGCTGCACCTGCCAGGGAACAAGTCCCCACACCGGGACCCTGCACCCCGAGGACCAGCTCG CT TCCTGCCACTACCAGGCCTGCCCCCCGCACTCCCGGAGCCACCCGGAATCCTGGCCCCCCAGCCCCCC GA TGTGGGCTCCTCGGACCCTCTGAGCATGGTGGGACCTTCCCAGGGCCGAAGCCCCAGCTACGCTTCCT GA (SEQ ID NO: 13) iCasp9 suicide gene ATGCTCGAGGGAGTGCAGGTGGAGACTATCTCCCCAGGAGACGGGCGCACCTTCCCCAAGCGCGGCCA GA CCTGCGTGGTGCACTACACCGGGATGCTTGAAGATGGAAAGAAAGTTGATTCCTCCCGGGACAGAAAC AA GCCCTTTAAGTTTATGCTAGGCAAGCAGGAGGTGATCCGAGGCTGGGAAGAAGGGGTTGCCCAGATGA GT GTGGGTCAGAGAGCCAAACTGACTATATCTCCAGATTATGCCTATGGTGCCACTGGGCACCCAGGCAT CA TCCCACCACATGCCACTCTCGTCTTCGATGTGGAGCTTCTAAAACTGGAATCTGGCGGTGGATCCGGA GT CGACGGATTTGGTGATGTCGGTGCTCTTGAGAGTTTGAGGGGAAATGCAGATTTGGCTTACATCCTGA GC ATGGAGCCCTGTGGCCACTGCCTCATTATCAACAATGTGAACTTCTGCCGTGAGTCCGGGCTCCGCAC CC GCACTGGCTCCAACATCGACTGTGAGAAGTTGCGGCGTCGCTTCTCCTCGCTGCATTTCATGGTGGAG GT GAAGGGCGACCTGACTGCCAAGAAAATGGTGCTGGCTTTGCTGGAGCTGGCGCGGCAGGACCACGGTG CT CTGGACTGCTGCGTGGTGGTCATTCTCTCTCACGGCTGTCAGGCCAGCCACCTGCAGTTCCCAGGGGC TG TCTACGGCACAGATGGATGCCCTGTGTCGGTCGAGAAGATTGTGAACATCTTCAATGGGACCAGCTGC CC CAGCCTGGGAGGGAAGCCCAAGCTCTTTTTCATCCAGGCCTGTGGTGGGGAGCAGAAAGACCATGGGT TT GAGGTGGCCTCCACTTCCCCTGAAGACGAGTCCCCTGGCAGTAACCCCGAGCCAGATGCCACCCCGTT CC AGGAAGGTTTGAGGACCTTCGACCAGCTGGACGCCATATCTAGTTTGCCCACACCCAGTGACATCTTT GT GTCCTACTCTACTTTCCCAGGTTTTGTTTCCTGGAGGGACCCCAAGAGTGGCTCCTGGTACGTTGAGA CC CTGGACGACATCTTTGAGCAGTGGGCTCACTCTGAAGACCTGCAGTCCCTCCTGCTTAGGGTCGCTAA TGCTGTTTCGGTGAAAGGGATTTATAAACAGATGCCTGGTTGCTTTAATTTCCTCCGGAAAAAACTTT TCTT TAAAACATCAGTCGACTATCCGTACGACGTACCAGACTACGCACTCGACTAA (SEQ ID NO: 14) RtTA Reverse tetracycline responsive transcriptional activator ATGTCTAGACTGGACAAGAGCAAAGTCATAAACGGCGCTCTGGAATTACTCAATGGAGTCGGTATCGA AG GCCTGACGACAAGGAAACTCGCTCAAAAGCTGGGAGTTGAGCAGCCTACCCTGTACTGGCACGTGAAG AA CAAGCGGGCCCTGCTCGATGCCCTGCCAATCGAGATGCTGGACAGGCATCATACCCACTTCTGCCCCC TG GAAGGCGAGTCATGGCAAGACTTTCTGCGGAACAACGCCAAGTCATTCCGCTGTGCTCTCCTCTCACA TC GCGACGGGGCTAAAGTGCATCTCGGCACCCGCCCAACAGAGAAACAGTACGAAACCCTGGAAAATCAG CT CGCGTTCCTGTGTCAGCAAGGCTTCTCCCTGGAGAACGCACTGTACGCTCTGTCCGCCGTGGGCCACT TT ACACTGGGCTGCGTATTGGAGGAACAGGAGCATCAAGTAGCAAAAGAGGAAAGAGAGACACCTACCAC CG ATTCTATGCCCCCACTTCTGAGACAAGCAATTGAGCTGTTCGACCGGCAGGGAGCCGAACCTGCCTTC CT TTTCGGCCTGGAACTAATCATATGTGGCCTGGAGAAACAGCTAAAGTGCGAAAGCGGCGGGCCGGCCG AC GCCCTTGACGATTTTGACTTAGACATGCTCCCAGCCGATGCCCTTGACGACTTTGACCTTGATATGCT GC CTGCTGACGCTCTTGACGATTTTGACCTTGACATGCTCCCCGGGTAA (SEQ ID NO: 15) Rabbit Beta-Globain PolyA signal (rBG) TCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACC AC TGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGG CT AATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACA TA TGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATA TG CTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCA TT CCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTT TC TTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCC CA GTCATAGCTGTCCCTCTTCTCTTATGGAGATC (SEQ ID NO: 16) UBC promoter GGTGCAGCGGCCTCCGCGCCGGGTTTTGGCGCCTCCCGCGGGCGCCCCCCTCCTCACGGCGAGCGCTG CC ACGTCAGACGAAGGGCGCAGCGAGCGTCCTGATCCTTCCGCCCGGACGCTCAGGACAGCGGCCCGCTG CT CATAAGACTCGGCCTTAGAACCCCAGTATCAGCAGAAGGACATTTTAGGACGGGACTTGGGTGACTCT AG GGCACTGGTTTTCTTTCCAGAGAGCGGAACAGGCGAGGAAAAGTAGTCCCTTCTCGGCGATTCTGCGG AG GGATCTCCGTGGGGCGGTGAACGCCGATGATTATATAAGGACGCGCCGGGTGTGGCACAGCTAGTTCC GT CGCAGCCGGGATTTGGGTCGCGGTTCTTGTTTGTGGATCGCTGTGATCGTCACTTGGTGAGTAGCGGG CT GCTGGGCTGGCCGGGGCTTTCGTGGCCGCCGGGCCGCTCGGTGGGACGGAAGCGTGTGGAGAGACCGC CA AGGGCTGTAGTCTGGGTCCGCGAGCAAGGTTGCCCTGAACTGGGGGTTGGGGGGAGCGCAGCAAAATG GC GGCTGTTCCCGAGTCTTGAATGGAAGACGCTTGTGAGGCGGGCTGTGAGGTCGTTGAAACAAGGTGGG GG GCATGGTGGGCGGCAAGAACCCAAGGTCTTGAGGCCTTCGCTAATGCGGGAAAGCTCTTATTCGGGTG AG ATGGGCTGGGGCACCATCTGGGGACCCTGACGTGAAGTTTGTCACTGACTGGAGAACTCGGTTTGTCG TC TGTTGCGGGGGCGGCAGTTATGGCGGTGCCGTTGGGCAGTGCACCCGTACCTTTGGGAGCGCGCGCCC TC GTCGTGTCGTGACGTCACCCGTTCTGTTGGCTTATAATGCAGGGTGGGGCCACCTGCCGGTAGGTGTG CG GTAGGCTTTTCTCCGTCGCAGGACGCAGGGTTCGGGCCTAGGGTAGGCTCTCCTGAATCGACAGGCGC CG GACCTCTGGTGAGGGGAGGGATAAGTGAGGCGTCAGTTTCTTTGGTCGGTTTTATGTACCTATCTTCT TA AGTAGCTGAAGCTCCGGTTTTGAACTATGCGCTCGGGGTTGGCGAGTGTGTTTTGTGAAGTTTTTTAG GC ACCTTTTGAAATGTAATCATTTGGGTCAATATGTAATTTTCAGTGTTAGACTAGTAAA (SEQ ID NO: 17) Example 5 A genetic cassette expressing a GLP-1, GIP, GCGR triple agonist is delivered using a lipid nanoparticle subcutaneously. The triple agonist is expressed under the regulation of a constitutive CMV promoter. The vector predominantly targets subcutaneous adipocytes and is injected into a specific locale subcutaneously into the abdomen lateral of the navel at a distance of approximately 3.0 cm. Prior to the injection of the therapy, a permanent tattoo is applied immediately to the right of the planned injection site (at a distance of 2-3mm away from the planned injection site). Following administration, medical records are updated indicating the location of the subcutaneous injection and position of the permanent tattoo in the form of a 0.5-1mm radius circle. If treatment cessation is desired, via for example ultrasound lipolysis, the lipolytic stimulus is applied to the site of injection and the permanent tattoo is used as a positioning guide. Similarly, should additional up-titration be required to augment the effect of the original treatment the tattoo can serve as a positioning guide enabling the application of a follow-up treatment at a different injection site. Should the treatment be stopped via for example application of ultrasound lipolysis and verification of the elimination of the therapeutic transgene from circulation (via serological biomarker analysis), the tattoo can be optionally removed using a tattoo removal laser. CMV Promoter TAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACAT AA CTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTA TG TTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCC CA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGC CC GCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGT CA TCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGG GG ATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTC CA AAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATAT AA GCAGAGCTGGTTTAGTGAACCGTCAGATC (SEQ ID NO: 18) Triple agonist sequence YAQGTFTSDYSILLDKKAQAAFIEYLLEGGPSSGAPPPS (SEQ ID NO: 19) Example 6 A dermal tattoo or a mark is administered at the time of treatment (such as during dosing or prior to dosing, but optionally up to 3 weeks following treatment) with the dose adjustable gene therapy, the tattoo or mark is administered at the site of treatment which includes a reporter element, which responds to substrate administration or other external stimulus (as an example irradiation by light or other non- ionizable radiation to excite a reporter construct). Following administration, the tattoo or mark remains visible at the site of injection allowing for dose adjustment to be performed locally at the site of injection. Such dose adjustment can be performed via pharmacological means, cryolipolysis, or ultrasound lipolysis. In addition, other means of evaluating the treatment such as injection site reaction scores, durability of expression, level of expression or other factors can be subsequently assessed at the time of injection. An illustration can be found in Figure 1. Example 7 A tattoo is administered to two subcutaneous injection sites on the left and right flank immediately prior to treatment or soon after initial treatment to create a permanent or semi permanent mark at the site of treatment administration. Said mark is subsequently used as a localizing element for the initial treatment to potentiate treatment of other areas, perform local dose adjustments, or evaluate the potency or durability of the initial signal. Dose adjustment is performed following treatment to reduce the level of gene expression using cryolipolysis, ultrasound lipolysis, or pharmacological dose adjustment via the administration of rapamycin locally at the site of injection in a 1mL injection. The marking remains following down-titration and can be used for any subsequent post treatment evaluation. An illustration can be found in Figure 2. Example 8 Markings were added to the site of injection immediately prior to treatment. The marking was used to locate the signal for subsequent quantification of gene expression using a reporter gene co-expressed with a therapeutic transgene. The expression was demonstrated to be correlated and both expression levels appeared to have a clear dose response curve. The reporter gene expression, which is quantified by a CCD camera is used to track the level of expression over time and ensure that the therapeutic benefit is afforded by the treatment in a minimally invasive manner. The CCD camera detects bioluminescent or fluorescent signals emitted from live tissue following substrate administration (luciferin or fluorofurimazine as an example). A CCD camera sensitive to low photone counts in the range of 1e5 to 1e10 photons per second is used to capture emitted light, allowing the analyst to non-invasively monitor transgene expression delivered by the system. An illustration can be found in Figure 3. Example 9: Subcutaneous Administration of GLP-1 Gene Therapy with Fluorescent Ink Marking and Cryolipolysis Down-Titration A patient is administered a subcutaneous injection of a non-viral gene therapy vector encoding the GLP-1 polypeptide. At the time of administration, the injection site is optionally marked with a biocompatible fluorescent ink. The marking is used as a spatial reference for follow-up treatment. At a subsequent time point, the marked area is visualized under appropriate light conditions and subjected to localized cryolipolysis to induce apoptosis in adipocytes at the injection site. This targeted depletion of transgene-expressing cells serves to reduce or eliminate expression of the GLP-1 transgene, thereby providing a means to down-titrate the therapeutic effect as clinically warranted. Example 10: Subcutaneous FGF21 Gene Therapy with Invisible Ink Marking and Bioluminescence Imaging of Reporter Expression A subcutaneous injection of a gene therapy vector encoding FGF21 is administered to a patient. The vector includes a co-expressed bioluminescent reporter gene, such as nanoluciferase. The injection site is optionally marked using an invisible ink that fluoresces under ultraviolet (UV) light, allowing for anatomical localization without visible marking in ambient conditions. At follow-up time points, bioluminescence imaging is performed using a CCD-based in vivo imaging system to non-invasively detect signal intensity from the reporter gene. This approach enables real-time monitoring of transgene expression dynamics. The invisible ink allows for consistent, site-specific imaging and optional delivery of follow-up treatments if necessary. Example 11: Subcutaneous Gene Therapy with Tattoo-Based Marking and Fluorescence-Based Expression Tracking A subcutaneous gene therapy is administered to a patient, encoding a GLP-1 / GIP dual agonist, optionally with a fluorescent reporter gene such as mCherry. The injection site is marked with a small, permanent tattoo to enable long-term anatomical reference. The tattoo marking facilitates clinical assessments of injection site reactions including erythema, edema, and lipodystrophy. In follow-up visits, the level of transgene expression is assessed using a fluorescence-based in vivo imaging system to detect the co-expressed reporter signal. This information is used to monitor persistence and spatial distribution of transgene expression over time and guide further clinical decision-making, including potential re-administration or targeted down-regulation interventions.

Claims

CLAIMS What is claimed is:

1. A system, which is a gene therapy treatment for subcutaneous administration, which contains elements that enable a physical, chemical, or biological viral or non-viral delivery vector, which is comprised of a) one or more elements that enable the system to be redosable following initial administration and enables predictable up- titration of gene expression b) one or more elements that enable a means of down-titration, to predictably reduce or optionally shut off the transgene expression c) and is co-formulated with elements or co-delivered with elements, at the time of treatment, that can serve as localization markers that enable post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration.

2. A physical element of delivering therapeutic transgenes in Claim 1, which includes one or more of electrical force, magnetic force, metallic or non-metallic particles accelerated traveling at a cell membrane penetrating velocity, physical force, or ultrasound.

3. A chemical element of delivering transgenes in Claim 1, which includes an organic, inorganic, or metalloorganic molecule which can act as a surfactant, cell penetrating agent, amphiphile, or a positively charged moiety to enable transport of genetic cargo to cells and to cellular nuclei.

4. A viral vector for delivering transgenes in Claim 1, which is a DNA, RNA, ssDNA, ssRNA, dsDNA, dsRNA, DNA-RNA hybrids, or a viral vector carrying a synthetic cargo.

5. A viral vector for delivering transgenes in Claim 1, which is an adeno-associated viral vector, adenoviral vector, lentiviral vector, retroviral vector, HSV, or a viral vector derived from a human, mammalian, or eukaryotic viral pathogen.

6. A viral vector for delivering transgenes in Claim 1, which is genetically, structurally, chemically or functionally modified, or optionally encapsulated, coated, or encapsidated into higherorder structures.

7. A viral vector for delivering transgenes in Claim 1, which is comprised at least in part of at least one known virus and at least one other element such as a targeting moiety, coating, formulation excipient, surfactant, lipid shell, or combination thereof.

8. A non-viral vector for delivering transgenes in Claim 1, which is comprised at least in part from a lipid nanoparticle formulation.

9. A non-viral vector for delivering transgenes in Claim 1, which is comprised at least in part from a polymeric nanoparticle formulation.

10. A non-viral vector for delivering transgenes in Claim 1, which is comprised at least in part from a lipid nanoparticle formulation with conjugated targeting moieties.

11. A non-viral vector for delivering transgenes in Claim 1, which is comprised at least in part from a polymeric nanoparticle formulation with conjugated targeting moieties.

12. A non-viral vector for delivering transgenes in Claim 1, which is comprised at least in part from a formulation of virus- like particles with or without conjugated targeting moieties.

13. An element of Claim 1 that enables redosability, which is a formulation that does not elicit a neutralizing antibody response.

14. An element of Claim 1 that enables redosability, which is a formulation that locally suppresses the immune response to enable the treatment without eliciting of a neutralizing antibody response.

15. An element of Claim 1 that enables redosability, which is a non-antigenic shell, capsule, or coating of the vector or genetic cargo.

16. An element of Claim 1 that enables redosability, which is an element enabling hypoimmune characteristics of the vector and / or genetic cargo.

17. An element of Claim 1 that enables down-titration, which is a genetic cassette delivered in cis, in trans, or encoded proximally to the therapeutic transgene that enables one or more of apoptosis of the carrier cell, a suppression of proteintranslation, a suppression of gene expression, epigenetic self- silencing, nuclease-mediated self-inactivation, or general shut- down of gene expression reducing the level of transgene expression from the therapeutic cassette.

18. An element of Claim 1 that enables down-titration, which is an element that targets a significant proportion of the genetic cargo to adipocytes, which can be eliminated by cryolipolysis, ultrasound lipolysis, liposuction, chemical induction of adipocyte apoptosis, or pharmacological induction of adipocyte apoptosis.

19. An element of Claim 1 that enables down-titration, which is a protein expressed from the delivered genetic cassette, that enables a transient or permanent reduction in the level of gene expression following stimulation by at least one internal or external stimulus.

20. A therapeutic transgene comprising the system in Claim 1, which encodes at least one human or mammalian protein or peptide.

21. A therapeutic transgene comprising the system in Claim 1, which encodes at least an antibody.

22. A therapeutic transgene comprising the system in Claim 1, which encodes at least a fusion protein.

23. A co-formulated element in Claim 1, which is a DNA sequence encoding a fluorescent protein or enzyme, which can be visualized in the fluorescent spectrum with or without inducible expression.

24. A co-formulated element in Claim 1, which is a DNA sequence encoding one or more of at least a part of Green Fluorescent Protein (GFP), enhanced GFP, enhanced GFP with a nuclear localization signal, superfolder GFP with S30R, Y39N, N105T, Y145F, I171V, and A206V variants, emergal green GFP, Venus, mNeon Green, EYFP, Ypet, Cerulean, CyPet, AmCyan, TagBFP, EBFP, dTomato, tdTomato, DsRed Express2, mRFP1, mApple, mKate, miRFP670nano, Turbo GFP, humanized recombinant GFP, destabilized GFP, , Yellow Fluorescent Protein, Cyan Fluorescent Protein, Red Fluorescent Protein, mCherry, TagRFP, mTurquoise, pHluorin2, supercliptic pH luorin, mt-mKeima, or a related protein analog, homolog, or paralog, or fragment thereof.

25. A co-formulated element in Claim 1, which is a DNA sequence encoding a pigment protein optionally under the regulation of an inducible promoter.

26. A co-formulated element in Claim 1, which is a DNA sequence encoding one or more of at least a fragment of bilirubin, hemocyanin, hemoglobin, myoglobin, reflectin, rhodopsin, myeloperoxidase, or other colored protein or encoding a set of DNA sequences enabling the production of pigments or pigment proteins.

27. A co-formulated element in Claim 1, which is a DNA sequence encoding a chemiluminescent reporter optionally under the regulation of an inducible promoter.

28. A co-formulated element in Claim 1, which is a DNA sequence encoding a reporter that is coupled with an inducer, optogenetic protein, or other regulatory element that enables its activation and visualization.

29. A co-formulated element in Claim 1, which is a DNA sequence encoding a chromogenic reporter optionally under the regulation of an inducible promoter.

30. A co-formulated element in Claim 1, which is a DNA sequence encoding one or more of at least a part of Firefly Luciferase, Renilla Luciferase, Gaussia Luciferase, NanoLuc Luciferase, Aequorin, Cypridina Luciferase, humanized firefly luciferase, Metridia Luciferase, Oplophorus Luciferase, Phrixothrix hirtus Luciferase, Renilla Reniformis Luciferase, or a related protein analog, homolog, or paralog, or fragment thereof.

31. A co-formulated element in Claim 1, which is a DNA sequence encoding a bioluminescent enzyme, which can be visualized following administration of an enzyme-specific substrate with or without inducible expression.

32. A co-formulated element in Claim 1, which is a pigment protein or peptide produced under the regulation of an inducible promoter.

33. A co-formulated element in Claim 1, which is a pigment protein or peptide produced under the regulation of a constitutive promoter.

34. A co-formulated element in Claim 1, which is an imaging contrast agent.

35. A co-formulated element in Claim 1, which is comprised of one or more nano- or micro-capsules, microspheres, lipid containing capsules or spheres, gadolinium based agent, metallic particles, a metal rod, sphere, or other geometric shape, a polymeric rod, sphere, or other geometric shape, a composite material rods, sphere, or other geometric shape.

36. A co-formulated element in Claim 1, which is comprised of one or more biocompatible dyes.

37. A co-formulated element in Claim 1, which is an adipocyte specific dye or a dye that is predominantly taken up by local subcutaneous adipocytes following administration.

38. A co-formulated element in Claim 1, which is a lipophilic biocompatible dye such as one or more of hematoxylin, eosin, oil red O, Congo red, alcain blue, rapid mucin, cresyl violet, Bielschowsky, fast blue, picrosirius red, Verhoeff van Gieson, Gomori’s trichrome, Masson’s trichrome, rapid PTAH, Prussian blue iron, AFB Kinyuon, AFB Ziehl-Neelson, Auramine O, Differential Quick, Fungi-Flour, Grocott Methenamine, TB Fluorostain, Warthin- Starry, Gram’s Stain, Von Kossa Dye, Villaneuva Osteochrome, Reticulin, or a biocompatible derivative or combination of biocompatible derivatives thereof.

39. A co-formulated element in Claim 1, which is comprised of one or more of carbon black, titanium dioxide, silver, or silver nitrates, copper phthalocyanine, logwood extract, cobalt blue, mercury sulfide, nickel, lead carbonate, or other metalloorganic, metallic, or organic agent that can be visualized directly or following application of high sensitivity photography, imaging, or enhancing electromagnetic radiation.

40. A co-formulated element in Claim 1, which is a die, protein, or contrast agent that is administered during the withdrawal of the administering needle to provide contrast to the dermis superficial to the site of injection, such as one or more of carbon black, titanium dioxide, copper phthalocyanine, logwood extract,cobalt blue, mercury sulfide, nickel, lead carbonate, or other metalloorganic, metallic, or organic agent that can be visualized directly or following application of high sensitivity photography, imaging, or enhancing electromagnetic radiation.

41. A co-delivered element in Claim 1, which is comprised of a patch, spray, label, polymer, or liquid that is applied at the time or prior to treatment and delivers a tattooing agent to mark the position of the treatment or up-titration.

42. A co-delivered element in Claim 1, which is a subcutaneously administrable marker that is opaque or partially opaque to ultrasound, or optionally palpable in the subcutis.

43. A co-delivered element in Claim 1, which is a subcutaneously administrable marker that is visible under ultraviolet light, or following irradiation by a specific range of electromagnetic waves.

44. A co-delivered element in Claim 1, which is an intradermally administrable marker that is opaque or partially opaque to ultrasound, or optionally palpable in the subcutis.

45. A co-delivered element in Claim 1, which is an intradermally administrable marker that is visible under ultraviolet light, or following irradiation by a specific range of electromagnetic waves.

46. A co-delivered element in Claim 1, which is capable of passively emitting radio waves or electromagnetic radiation following external stimulation.

47. A co-delivered element in Claim 1 which is capable of storing and / or processing and / or emitting information about the originally delivered gene therapy and optionally follow-up treatments such as up- and / or down-titrations.

48. A co-delivered element in Claim 1 capable of identifying the original location of the site of injection and optionally additional information via an RFID, Bluetooth low energy, near field communication, QR code, barcode, or other passive or active means.

49. A co-delivered element in Claim 1, which is a tattoo superficial to the administered gene therapy injection site.

50. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a chemical substance injected subcutaneously, intravenously, or ingested orally and enables bioluminescence of the reporter protein delivered in cis with the original therapeutic treatment.

51. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a luciferin, fluorofurimazine, coelenterazine, cypridina luciferin analog, hikarazine, aminoluciferin, d-luciferin salt, or related substance or combination thereof.

52. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a photoemitter with or without a coupled detected to detect and quantify fluorescence or bioluminescence from reporter protein markers delivered as part of the original gene therapy treatment.

53. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is an ultrasound system capable of detecting the co-formulated or co-administered marker.

54. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is one or more of a CT, MRI, SPECT, PET, infra-red, ultraviolet, or optical imaging system capable of detecting the co-formulated or co-administered marker with or without quantification.

55. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification,localization, quantification, evaluation, or targeting for up- and / or down-titration, which is a magnetic or electromagnetic emitter capable of detecting the co-formulated or co-delivered markers.

56. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is an RFID reader capable of interacting with one or more elements of the delivered marker.

57. An element used as part of the functionality of the system, but delivered at the time of post-treatment identification, localization, quantification, evaluation, or targeting for up- and / or down-titration, which is an electronic device capable of interacting with and optionally parsing the information encoded in the marker via scanning of a barcode or QR code, near field communication, Bluetooth low energy, wi-fi, or other electromagnetic or radiofrequency.

58. An element of enclosure of the system in Claim 1, which is a syringe, cartridge, vial, ampoule, or other primary container, which is optionally itself enclosed in a pen injector, autoinjector, safety syringe system, or patch injector.

59. A system described in the examples.

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