Telomerase upregulating polynucleotide and method of use and treatment thereof

Abstract

The present invention provides a composition comprising one or more polynucleotides capable of upregulating expression of telomerase wherein telomerase consists of a protein component with reverse transcriptase activity encoded by telomerase reverse transcriptase (TERT) gene and a non-coding RNA encoded by telomerase RNA component (TERC) gene, In an embodiment, the one or more polynucleotides of the present invention upregulates the telomerase by downregulating antisense RNA that regulates telomerase gene. In an embodiment, the one or more polynucleotides comprises an siRNA or an shRNA. In an embodiment, one or more polynucleotides of the present invention is delivered by a nanoparticle such as an exosome that encapsulates the one or more polynucleotides. The present invention also provides a method of treatment of telomerase deficiency related diseases or disorders using any embodiment of the one or more polynucleotides of the present invention.

Description

[0001] TELOMERASE UPREGULATING POLYNUCLEOTIDES AND METHOD OF USE AND TREATMENT THEREOF

[0002] INCORPORATION BY REFERENCE OF A SEQUENCE LISTING XML

[0003] A Sequence Listing is provided herewith as a Sequence Listing XML, “Telomerase Upregulating Polynucleotide Sequence LisLing.xml” created on January 15, 2025 and having a size of 132 KB. The contents of the Sequence Listing XML are incorporated by reference herein in their entirety.

[0004] FIELD OF THE INVENTION

[0005] The present invention provides a composition comprising one or more telomerase upregulating polynucleotides and method of use thereof.

[0006] BACKGROUND OF THE INVENTION

[0007] Telomerase is a ribonucleoprotein polymerase that maintains telomere ends by addition of the telomere repeat TTAGGG. Telomerase consists of a protein component with reverse transcriptase activity encoded by telomerase reverse transcriptase (TERT) gene and a non-coding RNA encoded by telomerase RNA component (TERC) gene that serves as a template for the telomere repeat. Telomerase expression plays a role in cellular senescence, as it is normally repressed in postnatal somatic cells resulting in progressive shortening of telomeres. Deregulation of telomerase expression in somatic cells may be involved in oncogenesis. Studies in mouse suggest that telomerase also participates in chromosomal repair, since de novo synthesis of telomere repeats may occur at double-stranded breaks. Increased telomerase expression has been found to play a role in cell senescence and aging and over-expression has been shown to ameliorate idiopathic pulmonary fibrosis (IFF) (https: / / doi.ore / 10.1083 / jcb.2020Q2120). Therefore, there is a need to upregulate expression of telomerase to treat lung diseases and potentially reversing fibrosis.

[0008] SUMMARY OF THE INVENTION

[0009] A pharmaceutical composition comprising a therapeutically effective amount of one or more telomerase upregulating polynucleotides, wherein the nucleotide sequence of each of the one or more telomerase upregulating polynucleotides is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 26, SEQ ID NO. 29, SEQ IDNO. 19, SEQ ID NO. 20, SEQ ID NO. 21, SEQ ID NO. 22, SEQ ID NO. 23, SEQ ID NO.24, SEQ ID NO. 25, SEQ ID NO. 17, SEQ ID NO. 27, SEQ ID NO. 28, SEQ ID NO. 18, SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 32, SEQ ID NO. 33, SEQ ID NO. 34, SEQ ID NO. 35, SEQ ID NO. 36, SEQ ID NO. 37, SEQ ID NO. 38, SEQ ID NO. 39, or a combination thereof.

[0010] A method of increasing the expression of TERT, TERC, or a combination thereof in a subject comprising the administration of a therapeutically effective amount of any embodiment of the one or more telomerase upregulating polynucleotides of the present invention to the subject.

[0011] A method of treatment of a telomerase-associated disease of a subject comprising the step of increasing the telomerase expression of the subject. In an embodiment, the step of increasing the telomerase expression of a subject of the present invention comprises the administration of a therapeutically effective amount of any embodiment of the one or more telomerase upregulating polynucleotides of the present invention to the subject .100012] BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 illustrates IncRNAs associated with the TERC locus. Top-candidate siRNA target sites for inhibiting the TERC-associated IncRNAs are also shown.

[0014] Figure 2 illustrates the IncRNAs associated with the TERT locus. Topcandidate siRNA target sites for inhibiting the TERT associated IncRNA ENST00000666708.1 which is a transcript of ENSG00000287486 are also shown.

[0015] Figure 3 illustrates the schematic showing the siRNAs targeted to human TERC-associated IncRNAs and used in the screening of the human TERC-associated IncRNAs.

[0016] Figure 4 illustrates the screening of siRNAs that target TERC-associated IncRNAs. TERC expression was assessed in HEK293 siRNA (lOOnM) transfected cells. RNA was collected 72hrs post-transfection and qRT-PCR carried out for TERC expression. The averages of triplicated treated cultures are shown with the standard deviations and P values from a paired T-test (two-tailed si2 and one-tailed silO).

[0017] Figure 5 illustrates the effects of various siRNAs on TERC expression. HEK293 siRNA (lOOnM) transfected cells were collected 72hrs post-transfection and qRT- PCR carried out for TERC expression. The averages of triplicated treated cultures are shown with the standard deviations and P values from a paired T-test.

[0018] Figure 6 illustrates the top candidate siRNAs for activation of TERC. Several siRNAs were screened targeting TERC-associated antisense RNAs. The top candidates and the TERCF1 / R1 or TERCF2 / R2 amplicons used in the qRT-PCR assessment are shown relative to the TERC gene.

[0019] Figure 7 illustrates the siRNA target sites in 3* TERT associated antisense IncRNA (ENSG00000287486).

[0020] Figure 8 illustrates the siRNA-mediated activation of TERT in HEK293 cells. HEK293 cells were transfected (lOOnM, L2K) and assessed TERT mRNA expression 72hrs later.

[0021] Figure 9 illustrates the siRNA-mediated activation of TERT in HEK293 cells. HEK293 cells were transfected with the indicated siRNA for 72 hr and harvested (12-well plate). RNA was extracted using RNeasy kit with DNase treatment and qPCR primer beta actin; TERT Fl, R1 (Table 1).

[0022] Figure 10 illustrates the pooled expression of TERT mRNAs in E666-si4-l and si4-2 treated cells. The averages of triplicate treated HEK cells are shown with the standard deviations.

[0023] Figure 11 illustrates the vector map of the dual expressing shRNA EV producing vector. The vector pRP[Exp]-{pshTERC10&13_Dual]-CMV>Puro was cotransfected with the shRNA packaging and enhancer vector from the RNex system (patent applications No. 63 / 598499 and 63 / 607,552) and cells multiple drag selected.

[0024] Figure 12 illustrates the schematic depicting the transwell experimental approach used to evaluate the packaging and transfer of therapeutic RNAs using the RNex system for functional efficacy. Producer cells are transfected with combinations of plasmids which reprogram these cells to generate extracellular vessicles (EVs) packaged with therapeutic RNAs targeted to various diseases. The EV-asRNAs, EV-shRNAs or EV- mRNAs, generated from the reprogrammed producer cells permeate the 0.4pM membranes and can be taken up by the recipient cells. The recipient cells can be stable reporter transduced cell lines or HEK293 cells directly transfected with various reporter plasmids containing the therapeutic RNA target site in fusion with various reporter transgenes, RLuc and GFP or patient derived organoid cells and the effects on the targeted disease relevant gene determined.

[0025] Figure 13 illustrates the activation of TERC using the dual expression shRNA EV producing vector of Figure 11 (shTERC-2+shTERC- 10). Drag selected line 2 producer cells were co-cultured with recipient HEK293 cells using a transwell (Figure 12). Therecipient cells were collected following 4 days (96hrs) of co-culturing in replicates of n=3 and assessed by qRT-PCR for the particular target.

[0026] DETAILED DESCRIPTION OF THE INVENTION

[0027] As used in this specification and in claims which follow, the singular forms “a”, “an” and “the” include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to “an ingredient” includes mixtures of ingredients, reference to “an active pharmaceutical agent” includes more than one active pharmaceutical agent, and the like.

[0028] As used herein, the term “about” as a modifier to a quantity is intended to mean + or - 5%%, + or - 10%, + or - 15% or + or - 20% inclusive of the quantity being modified.

[0029] As used herein, the term "nucleic acid" refers to nucleotides (e.g., deoxyribonucleotides or ribonucleotides) and polymers thereof in either single-, double- or multiple-stranded form, or complements thereof. The terms "polynucleotide," "oligonucleotide," "oligo" or the like refer, in the usual and customary sense, to a linear sequence of nucleotides. The term "nucleotide" refers, in the usual and customary sense, to a single unit of a polynucleotide, i.e., a monomer. Nucleotides can be ribonucleotides, deoxyribonucleotides, or modified versions thereof. Examples of nucleic acids contemplated herein include single and double stranded DNA, single and double stranded RNA, and hybrid molecules having mixtures of single and double stranded DNA and RNA. Examples of nucleic acids contemplated herein include any types of RNA (e.g., antisense RNA, mRNA, siRNA, miRNA, shRNA, guide RNA, dicer substrate RNA, dicer substrate siRNAs (dsiRNAs) (dsiRNA are cleaved by the RNase I class endoribonuclease dicer into 21-23 base duplexes having 2-base 3'-overhangs siRNA), and any type of DNA, genomic DNA, plasmid DNA, and minicircle DNA, and any fragments thereof. The term "duplex" in the context of nucleic acids refers, in the usual and customary sense, to double strandedness. Nucleic acids can be linear or branched. For example, nucleic acids can be a linear chain of nucleotides or the nucleic acids can be branched, e.g., such that the nucleic acids comprise one or more arms or branches of nucleotides. Optionally, the branched nucleic acids are repetitively branched to form higher ordered structures such as dendrimers and the like, In an embodiment, the nucleotide sequence is provided using symbols ATCG (adenine (A), cytosine (C), guanine (G), and thymine (T)) for a DNA molecule, and provided with codes using symbols AUCG (adenine (A), cytosine (C), guanine (G), and uracil (U)) for a RNA molecule. In anembodiment, the symbols T and U are used interchangeably in a nucleotide sequence to illustrate the DNA and RNA molecule respectively made according to the nucleotide sequence.

[0030] As used herein, the terms "polypeptide," "peptide" and "protein" generally refer to a polymer of amino acid residues. As used herein, the term also applies to amino acid polymers in which one or more amino acids are chemical analogs or modified derivatives of corresponding naturally occurring amino acids or are unnatural amino acids. The term "protein", as generally used herein, refers to a polymer of amino acids linked to each other by peptide bonds to form a polypeptide for which the chain length is sufficient to produce tertiary and / or quaternary structure. In an embodiment, the "polypeptide," "peptide" or "protein" of the present invention is prepared from a plasmid encoding said "polypeptide," "peptide" or "protein". Therefore, the "polypeptide," "peptide" or "protein" of the present invention further comprises a nucleotide sequence encoding said "polypeptide," "peptide" or "protein" that could be converted using a genetic code such as but not limited to the standard genetic code.

[0031] As used herein, “sequence identity” and “% identity,” refers to the value determined by comparing two optimally aligned sequences over a comparison window, wherein a portion of the sequence in the comparison window may comprise additions or deletions as compared to the reference sequence for optimal alignment of the two sequences. The number of positions at which identical amino acid residues occur in both sequences is determined, yielding the number of matched positions, which is divided by the total number of positions in the window of comparison and the result multiplied by 100 to yield the percentage of sequence identity. The comparison window is the entire length of the sequence being referred to unless indicated otherwise.

[0032] As used herein, “% similarity” is calculated as described for “% identity,” with the exception that the hydrophobic residues Ala, Vai, Phe, Pro, Leu, De, Tip, Met, and Cys are similar, the basic residues Lys, Arg, and His are similar; the acidic residues Glu and Asp are similar, and the hydrophilic, uncharged residues Gin, Asn, Ser, Thr, and Tyr are similar. The remaining natural amino acid Gly is not similar to any other amino acid in this context.

[0033] As used herein, the term “subject,” “individual” or “patient” is used interchangeably herein, which refers to a vertebrate, preferably a mammal, more preferably a human. Mammals include, but are not limited to, murines, simians, humans, farm animals, sport animals, and pets.

[0034]

[0035] As used herein, the term “effective amount” or “a therapeutically effective amount” of a drug, compound, pharmacologically active agent or a pharmaceutical composition comprises administering an amount thereof necessary to achieve a desired result. The exact amount required will vary from subject to subject, depending on the species, age, general condition of the subject, the severity of the disease, the particular active agent, its mode of administration, the desired outcome, and the like. In certain embodiments of the present invention, a “therapeutically effective amount” of a drug, compound, pharmacologically active agent or a pharmaceutical composition is that amount effective for inhibiting progression or reversing of any disease disclosed herein in a subject or a biological sample (e.g., in cells). In certain embodiments, disease progression is inhibited by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 100%. In certain embodiments, the drug, compound, pharmacologically active agent or a pharmaceutical composition inhibits disease progression by at least about 25%, at least about 50%, at least about 75%, at least about 90% or at least about 100%. In certain embodiments of the present invention, a “therapeutically effective amount” refers to an amount of a drug, compound, pharmacologically active agent or a pharmaceutical composition sufficient to cause reversal of disease. In certain embodiments, the disease is reversed by about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 100% or any numbers and number ranges falling within these values.

[0036] As used herein, the term “target” or “targeting” a polynucleotide comprises directly or indirectly regulating the expression level, biological function, or a combination thereof, of said polynucleotide. The indirect regulation of a polynucleotide comprises indirectly regulating the expression level, biological function, or a combination thereof, of a polynucleotide by regulation of one or more antisense RNA that regulates the polynucleotide being targeted. In an embodiment, in the case of direct regulation, down regulating a polynucleotide may comprise destruction or breakup of the polynucleotide being targeted or regulated. In an embodiment, in the case of indirect regulation, down regulating a polynucleotide may comprise destruction or breakup of one or more antisense RNA that regulates the polynucleotide being targeted. In an embodiment, such destraction or breakup is done using a small interfering RNA (siRNA), a short hairpin RNA (shRNA), a microRNA (miRNA), a ribozyme, a deoxyribozyme, an aptamer, or a combination thereof targeting or regulating the polynucleotide.

[0037] The present invention provides one or more telomerase upregulating polynucleotides and a pharmaceutical composition thereof. In an embedment, the one or more telomerase upregulating polynucleotides of the present invention regulates telomerase expression through regulating the expression level of TERT, TERC, or a combination thereof. In an embodiment, the regulation of the expression level of TERT, TERC, or a combination thereof comprises the regulation of the expression level of an antisense regulator of TERT, TERC, or a combination thereof. In an embodiment, the one or more telomerase upregulating polynucleotides targeting TERT, TERC, or a combination thereof of the present invention increases or de-represses the expression level of TERT, TERC, or a combination thereof by suppressing the expression level of an antisense RNA regulator of TERT, TERC, or a combination thereof. In an embodiment, the antisense RNA regulator of TERT, TERC, or a combination thereof is antisense to the promoter of TERT, TERC, or a combination thereof. In an embodiment, the nucleotide sequence of the antisense RNA regulator of TERT, TERC, or a combination thereof is at least about 80%, about 85%, about 90%, about 95%, or about 100% identical to the nucleotide sequence of AW207347 (SEQ ID NO. 1), AIS 25849 (SEQ ID NO. 2), AW136367 (SEQ ID NO. 3), A1380754 (SEQ ID NO. 4), AW293800 (SEQ ID NO. 5), BM695101 (SEQ ID NO. 6), CV370609 (SEQ ID NO. 7), MG677549 (SEQ ID NO. 8), BF802688 (SEQ ID NO. 9), AA748707 (SEQ ID NO. 10), AA811084 (SEQ ID NO. 11), AI824948 (SEQ ID NO. 12), AW270031 (SEQ ID NO. 13), AW276315 (SEQ ID NO. 14), DB345416 (SEQ ID NO. 15) deposited in the NCBI Nucleotide database (https: / / www.ncbi.nlm.nih.gov / nuccore / ) or ENST00000000000666708.1 (SEQ ID NO. 16) deposited in the Ensembl database (https: / / asia.ensembl.org / index.html) of a subject. In an embodiment, the subject comprises a human cell.

[0038] SEQ ID NO. 1 (AW207347):TTTTTTTTTTTTTTTTTCAGGTTTGGGGGTTCACAAGCCCCCATTGCCGGCGAGGGGTGACGGATGCGCACGATCGGCGTTCCCCCCACCAACAGGAAAGCGAACTGCATGTGTGAGCCGAGTCCTGGGTGCACGTCCCACAGCTCAGGGAATCGCGCCGCGCGCGGGGACTCGCTCCGTTCCTCTTCCTGCGGCCTGAAAGGCCTGAACCTCGCCCTCGCCCCCGAGAGACCCGCGGCTGACAGAGCCCAACTCTTCGCGGTGGCAGTGGGTGCCTCCGGAGAAGCCCCGGGCCGACCGCGGGCTCCAGGCGGGGTTCGGGGGCTGGGCAGGCGACCCGCCGCAGGTCCCCGGGAGGGGCGAACGGGCCAGCAGCTGACAl l'l'l l lG

[0039] SEQ ID NO. 2 (AI825849):AGGTTTGGGGGTTCACAAGCCCCCATTGCCGGCGAGGGGTGACGGATGCTCACGATCGGCGTTCCCCCCACCAACAGGAAAGCGAACTGCATGTGTGAGCCGAGTCCTGGGTGCACGTCCCACAGCTCAGGGAATCGCGCCGCGCGCGGGGACTCGCTCCGTTCCTCTTCCTGCGGCCTGAAAGGCCTGAACCTCGCCCTCGCCCCCGAGAGACCCGCGGCTGACAGAGCCCAACTCTTCGCGGTGGCAGTGGGTGCCTCCGGAGAAGCCCCGGGCCGACCGCGGCCTCCAGGCGGNGTTCGGNGGCTGGGCAGGCGACCCGCCGCAGGTCCCCGGGAGGGGCGAACGGGCCAGCAGCTGACATmTTGTTTGCTCTAGAATGAACGGTGGAAGGCGGCAGGCCGAGGCTTTTCCGCCCGCTGAAAGTCAGCGAGAAAAACA

[0040] SEQ ID NO. 3 (AW136367):TrTTTTTTTTTTTTTTTACTGCATGTGTGAGCCGAGTCCTGGGTGCACGTCCCACAGCTCAGGGAATCGCGCCGCGCGCGGGGACTCGCTCCGTTCCTCTTCCTGCGGCCTGAAAGGCCTGAACCTCGCCCTCGCCCCCGAGAGACCCGCGGCTGACAGAGCCCAACTCTTCGCGGTGGCAGTGGGTGCCTCCGGAGAAGCCCCGGGCCGACCGCGGCCTCCAGGCGGGGTTCGGGGGCTGGGCAGGCGACCCGCCGCAGGTCCCCGGGAGGGGCGAACGGGCCAGCAGCTGACATTTTTTGTTTGCTCTAGAATGAACGGTGGAAGGCGGCAGGCCGAGGCTTTTCCGCCCGCTGAAAGTCAGCGAGAAAAACAGCGCGCGGGGAGCAAAAGCACGGCGCCCCTCGTGCCG

[0041] SEQ ID NO. 4 (AI380754):TnTCTTCTGCATGTGTGAGCCGAGTCCTGGGTGCACGTCCCACAGCTCAGGGAATCGCGCCGCGCGCGGGGACTCGCTCCGTTCCTCTTCCTGCGGCCTGAAAGGCCTGAACCTCGCCCTCGCCCCCGAGAGACCCGCGGCTGACAGAGCCCAACTCTTCGCGGTGGCAGTGGGTGCCTCCGGAGAAGCCCCGGGCCGACCGCGGCCTCCAGGCGGGGTTCGGGGGCTGGGCAGGCGACCCGCCGCAGGTCCCCGGGAGGGGCGAACGGGCCAGCAGCTGACATTTTTTGTTTGCTCTAGAATGAACGGTGGAAGGCGGCAGGCCGAGGCTTTTCCGCCCGCTGAAAGTCAGCGAGAAAAACAGCGCGCGGGGAG

[0042] SEQ ID NO. 5 (AW293800):TITITrCCTTTTTTTCTGCATGTGTGAGCCGAGTCCTGGGTGCACGTCCCACAGCTCAGGGAATCGCGCCGCGCGCGGGGACTCGCTCCGTTCCTCTTCCTGCGGCCTGAAAGGCCTGAACCTCGCCCTCGCCCCCGAGAGACCCGCGGCTGACAGAGCCCAACTCTTCGCGGTGGCAGTGGGTGCCTCCGGAGAAGCCCCGGGCCGACCGCGGCCTCCAGGCGGGGTTCGGGGGCTGGGCAGGCGACCCGCCGCAGGTCCCCGGGAGGGGCGAACGGGCCAGCAGCTGACATnTTTGTTTGCTCTAGAATGAACGGTGGAAGGCGGCAGGCCGAGGCTTTTCCGCCCGCTGAAAGTCAGCGAGAAAAACAGCGCGCGGGGAGCAAAAGCACGGCGCCTACGCCCTTCTCAGTTAGGGGT

[0043] SEQ ID NO. 6 (BM695101):TGACCATTAAAGGAACACAATTTCCAATGTTCATTTAGATCTTCTAATTAAATATTCATTAAATGTTAAATGATCTCTCAAAAAAAAATGACTGTTCTCCCACACCCCGTTGAGGGGACTGGTCGAGATCTACCTTGGGAGAAGCAAAAACCTCAACAAAATCTGCAGAGCAGGAACTAAGTTGTAATACAACCATAAAAGGCAACAAAAAGCGGAAGACGGGAGAACCCACGCAGGAACGGCTCCAGGCAACCCCGGCTCACTGCCCATTCATTTTGGCCGACnTGGAGGTGCCTTCACGTCTCCTGCCAATTTGCAGCACACTGGCCCAGTCAGTCAGGTTTGGGGGCTC

[0044] SEQ ID NO. 7 (CV370609):TnTCCGCCCGCTGAAAGTCAGCGAGAAAAACAGCGCGCGGGAAGCAAAAGCACGGTGCCTACGCACTTCTCAGTTAGGGTTAGACAAAAAATGGNCACCAACCCTCCCAGGGCCAC

[0045] SEQ ID NO. 8 (MG677549):AGTCCCTCCGCCACGTGGGAAGCGCGGTCCTGGGCGTCTGTGCCCGCGAATCCACTGGGAGCCCGGCCTGGCCCCGACAGCGCAGCTGCTCCGGGCGGACCCGGGGGTCTGGGCCGCGCTTCCCCGCCCGCGCGCCGCTCGCGCTCCCAGGGTGCAGGGACGCCAGCGAGGGCCCCAGCGGAGAGAGGTCGAATCGGCCTAGGCTGTGGGGTAACCCGAGGGAGGGGCCATGATGTGGAGGCCCTGGGAACAGGTGCGTGCGGCGACCCTTTGGCCGCTGGCCTGATCCGGAGACCCAGGGCTGCCTCCAGGTCCGGACGCGGGGCGTCGGGCTCCGGGCACCACGAATGCCGGACGTGAAGGGGAGGACGGAGGCGCGTAGACGCGGCTGGGGACGAACCCGAGGACGCATTGCTCCCTGGACGGGCACGCGGGACCTCCCGGAGTGCCTCCCTGCAACACTTCCCCGCGACTTGGGCTCCTTGACACAGGCCCGTCATTTCTCTTTGCAGGTTCTCAGGCGGCGAGGGGTCCCCACCATGAGCAAACCACCCCAAATCTGTTAATCACCCACCGGGGCGGTCCCGTCGAGAAAGGGTGGGAAATGGAGCCAGGCGCTCCTGCTGGCCGCGCACCGGGCGCCTCACACCAGCCACAACGGCCTTGACCCTGGGCCCCGGCACTCTGTCTGGCAGATGAGGCCAACATCTGGTCACATCCCGCCCGCACAGGGTGGAGGGCAACCTCGGGGTCCAGGCACCTGGCTCCAAGCCTCGGACTGCAGAGCTAGGAGGCCCGACTTCCAGCCCAGCAGTAGAAGCCACACGGCCACTGGTCCCCTCCAGACCTGGGGCCCCGGCACAACCGCAGGACAGCTGAGGACTTCCCAGGAATCCAGACTCCGGGTTGCTCAAGTTTGGATCTAAGGGGCGAGAAACTTCTGGGTCTCCCGAGGCCTTGCAGGGATGCTGTAGCTGAGGTCGGCAAACACTGAAATGCTAACAAACGCAACCTTAAATGTAACCTTTCCTACTTTCAGAAACTGCCGGAGGAAATTGCTTTATTTATGGAGCTAGCATTTGAACAGGCCTCGCACCCTCCCTGGGCTGTCACGCTCGCTGGAGGTTAGCCTCGTCTTGTAAATACTTAGGATTACAGGTCGCTCTTCTAGAAATCCCCTTAGTGATCCCTAAGCCTTTTTAAAGGGCTGTGTTTGTGAATTGTCTCTGCCACTAGGGCAAAGGGGCGGTrTGGAAAATTTGTTCCAACAAAAGTTAAGTTGTAGCTrACACTGGTTCTCTGCAGAGAAGCCAACATAGAAAACACAATTTTAAAAGAGGGAAGAGAAGAAATGGAAGCAGAAGATTATGCTGGAGTAATTAACACCATGTGCATGGCGAGGAAACGCCTCCCGGCATTCAATGAAGATCGCTGATACCCAGAAGACACCCCAGTATTATGGGTGCAGTTAGTGTGTCTTTGAAAAGCTGATGATGTCTTAGTCATCACAGTGTAAAACATCAAGAGTGTTCTAACAACAATAAAAAAATTCTATCATTGGCTTAAAACACCACAACACTTGAGTGGGGTGAGCTTCCTACCTCAGACCCAGATGTTrC

[0046] SEQ ID NO. 9 (BF802688):GATTAGAGGATCCCAGTAGTCGGACGTGTAGTGAATGAGATGATGGTCTCACACCATCACCCAGACCAGGTTATGCACACAGGGCGGTGGTCAGAAGGATTGGCAGGACGTTGATATACGATGACATCAGGTTGTCTGACGAAGGCAGGATTCATGATAAGTACTCTrGACGCTGTCCGCATCCTCTCAGGTTCACGCATGTGTGCTGCAGCTCCCATTTCATCAGCAAGTTTGGAAGAACCCCACATTTTTCCTGCGCGTCATCTCTGACACGGCCTCCCTCTGCTACTCCATCCTGCGGTCCTGAGCTTAACAGCTTCTACTTTCTGTTCTTTCTGTGTTGTGGAAATTTCACCTCCCAGGGCGGGTGCCAGGCTCGCAGTGGAGCTGGACATACGTCCTTCCTCAGGCAGAAGGAACTGGAAGGATTGCAGAGAACAGGAGGGGCGGCTCAGAGGGACGCAGTCTTGGGGTGAAGAAACAGCCCCTCCTCAGAAGTTGGCTTGGGCCACACGCGGTC

[0047] SEQ ID NO. 10 (AA748707):TTTTTTTCAAAACTGAAAAACTCATATATTCAGTATTTTACTCCCACAGCACCTCCCCCCAATTTGACCCACAGGGACCCCCATCCAGGTGCAGGGTCCTCGCCTGTGTACAGGGCACACCTTTGGTCACTCCAAATTCCCAGAGCTCCCAGGGTCCTTCTCAGGGTCTCCACCTGGATGGTGGGCCCCGGCCCCCAAACCACGGCACGGCGAGGGGTGAACAATGGCGAATCTGGGGATGGACTATTCCTATGTGGGGAGTGGAAGCCGGGCTCCTGGTGAGGAAAAGCTGGCCCTGGGGTGGAGCCGAGCGCA

[0048] SEQ ID NO. 11 (AA811084):TTTTTTTTTTTTTTTTTTCAAAACTGAAAAACTCATATATTCAGTATTTTACTCCCACAGCACCTCCCCCCAATTTGACCCACAGGACCCCCATCCAGGTGCAGGGTCCTCGCCTGTGTACAGGGCACACCTTTGGTCACTCCAAATTCCCAGAGCTCCCAGGGTCCTTCTCAGGGTCTCCACCTGGATGGTTGGGGTGGAAGGCAAAGGAGGGCAGGGCGAGGGGTGAACAATGGCGAATCTGGGGATGGACTATTCCTATGTGGGGAGTGGAAGCCGGGCTCCTGGTGAGGAAAAGCTGGCCCTGGGGTGGAGCCGAGCGCCACCTGTGGGGAAGTGA

[0049] SEQ ID NO. 12 (AI824948):TTTTITITnTTTTTCAAAACTGAAAAACTCATATATTCAGTATTTrACTCCCACAGCACCTCCCCCCAATTTGACCCACAGGGACCCCCATCCAGGTGCAGGGTCCTCGCCTGTGTACAGGGCACACCTTTGGTCACTCCAAATTCC

[0050] SEQ ID NO. 13 (AW270031):TTCAAAACTGAAAAACTCATATATTCAGTATTTTACTCCCACAGCACCTCCCCCCAATTTGACCCACAGGGACCCCCATCCAGGTGCAGGGTCCTCGCCTGTGTACAGGGCACACCTTTGGTCACTCCAAATTCCCAGAGCTCCCAGGGTCCTTCTCAGGGTCTCCACCTGGATGGTGGGGGTGGAAGGCAAAGGAGGGCAGGGCGAGGGGTGAACAATGGCGAATCTGGGGATGGACTATTCCTATGTGGGGAGTGGAAGCCGGGCTCCTGGTGAGGAAAAGCTGGCCCTGGGGTGGAGCCGAGCGCCAGCCTGTGGNGAAGTGAAGACGGCAGGTGTGCTGGACACTCAGCCCTTGGCTGGACACTCGCTCAGGCCTCAGCCGGACACTCAGCCTTCAACCGGACATGCAGGCCTCGGCCAAACACTCACTCAGGCCTCAGACTCCCAGCGGTGCGGGCCTGGGTG

[0051] SEQ ID NO. 14 (AW276315):TCAAAACTGAAAAACTCATATATTCAGTATTTTACTCCCACAGCACCTCCCCCCAATTTGACCCACAGGGACCCCCATCCAGGTGCAGGGTCCTCGCCTGTGTACAGGGCACACCTTTGGTCACTCCAAATTCCCAGAGCTCCCAGGGTCCTTCTCAGGGTCTCCACCTGGATGGTGGGGGTGGAAGGCAAAGGAGGGCAGGGCGAGGGGTGAACAATGGCGAATCTGGGGATGGACTATTCCTATGTGGGGAGTGGAAGCCGGGCTCCTGGTGAGGAAAAGCTGGCCCTGGGGTGGAGCCGAGCGCCAGCCTGTGGGGAAGTGAAGACGGCAGGTGTGCTGGACACTCAGCCCTTGGCTGGACACTCGCTCAGGCCTCAGCCGGACACTCAGCCTTCAGCCGGACATGCAGGCCT

[0052] SEQ ID NO. 15 (DB345416):CAAAACTGAAAAACTCATATATTCAGTATTTTACTCCCACAGCACCTCCCCCCAATTTGACCCACAGGGACCCCCATCCAGGTGCAGGGTCCTCGCCTGTGTACAGGGCACACCTTTGGTCACTCCAAATTCCCAGAGCTCCCAGGGTCCTTCTCAGGGTCTCCACCTGGATGGTGGGGGTGGAAGGCAAAGGAGGGCAGGGCGAGGGGTGAACAATGGCGAATCTGGGGATGGACTATTCCTATGTGGGGAGTGGAAGCCGGGCTCCTGGTGAGGAAAAGCTGGCCCTGGGGTGGAGCCGAGCGCCAGCCTGTGGGGAAGTGAAGACGGCAGGTGTGCTGGACACTCAGCCCTTGGCTGGACACTCGCTCAGGCCTCAGCCGGACACTCAGCCTTCAGCCGGACATGCAGGCCTCGGCCAAACACTCACTCAGGCCTCANACTCCCAGCGGTGCGGGCCTGGGTGTGGGCCGCCCCTCCCTCCCTGGGACGTANAGCCCGGCGTGACAGGGCTGCTGGTGTCTGCTCTCGGCCTGGCT

[0053] SEQ ID NO. 16 (ENST 566708.1):CACGTCCGGGCTGCACTCAAACCAGAAGCCCTGTCCAAGGTGCACTCAGTAGGACCCGGCCATACCATAAGGCCCCACCAGGGACCTGCTGGAAGGCTCTGGAGCCACTCACGCAGGGCCCTCGGTTTCGTGTGGCCCAAGCCAACTTCTGAGGAGGGGCTGTTTCTTCACCCCAAGACTGCGTCCCTCTGAGCCGCCCCTCCTGTTCTCTGCAATCCTTCCAGTTCCTTCTGCCTGAGGAAGGACGTATGTCCAGCTCCACTGCGAGCCTGGCACCCGCCCTGGGAGGTGAAATTTCCACAACACAGAAAGAACAGAAAGTAGAAGCTGTTAAGCTCAGGACCGCAGGATGGAGTAGCAGAGGGAGGCCGTGTCAGAGATGACGCGCAGGAAAAATGTGGGGTTCTTCCAAACTTGCTGATGAAATGGGAGCTGCAGCACACATGCGTGAAACCTGAGAGGATGGCGGACAGCGTCAGAGGAAAGGCCTCCTAATCAGACGGTGCTCGTGGGTGTGGGCATGGGCCCACCGGTGCCTGTGTGCGTGCATGAATGCACATGCATGGGTTTCCTCATGGGCACAGGTGCACACACACGGATGCATGCATGCATGTCTGTGTGTGTGCTTGTGTGTGCGAGTAGCTGCGTGTCTGTGTGTGCACAGGTACACTGCGTTTCTGTGCACCATCTGTATGAACACGCATGTGGAGGCTGAAGCAGCTCCATCCTGGATGCCAGCCCCCCATGCTGGCTTCTGATTAGCCCCTGTTCCGGGAAGGCCTCTAAGGTTTCCAGTTTATCCATTGTTCCTTGTGTACCAGCAGGTACTTATCATGAATCCTGCCCTTCGTCAGACAACCTTGATGTCATCGTATATCAACGTCCTGCGCATCCCTTCTGAGCCACCCGCCCCTGTGGTGCATAAACCCTGGGTCTGGGGTGATGGTGTGAGGACCCATCATCTCATCTCACTGACACCGAGACACCGACTTGGCTTCTGTGTGTAGGTTAAGTTCCTA

[0054] In an embodiment, the telomerase upregulating polynucleotides of the present invention is about 10 to about 100 nucleotides (nt) in length such as about 10, about 20, about30, about 40 about 50 about 60 about 70, about 80, about 90, or about 100 nt in length including any lengths or length ranges falling within these values. In an embodiment, the telomerase upregulating polynucleotides of the present invention is about 15 to about 55 nt in length, about 20 to about 30 nt in length or about 21 to about 23 nt in length. In an embodiment, the telomerase upregulating polynucleotides of the present invention is double stranded or single stranded. In an embodiment, the telomerase upregulating polynucleotides of the present invention is blunt ended or comprise overhanging ends. In an embodiment, the telomerase upregulating polynucleotides of the present invention is chemically synthesized or recombinantly produced. In an embodiment, one strand of the telomerase upregulating polynucleotides of the present invention comprises nucleotide sequence having sufficientcomplementarity to the antisense RNA regulator of TERT, TERC, or a combination thereof for the polynucleotide to direct cleavage of the said antisense RNA regulator via RNA interference.

[0055] In an embodiment, the one or more telomerase upregulating polynucleotides of the present invention comprise a small interfering RNA (siRNA), a short hairpin RNA (shRNA), a microRNA (miRNA), a ribozyme, a deoxyribozyme, an aptamer, or a combination thereof. In an embodiment, the one or more telomerase upregulating polynucleotides of the present invention comprise siRNA. In an embodiment, nucleotide sequence of the siRNA telomerase upregulating polynucleotides of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to the nucleotide sequence of SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 19, SEQ ID NO. 20, SEQ ID NO. 21, SEQ ID NO. 22, SEQ ID NO. 23, SEQ ID NO. 24, SEQ ID NO. 25, SEQ ID NO. 26, SEQ ID NO. 27, SEQ ID NO. 28, SEQ ID NO. 29, SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 32, SEQ ID NO. 33, SEQ ID NO. 34, SEQ ID NO. 35, SEQ ID NO. 36, SEQ ID NO. 37, SEQ ID NO. 38, or SEQ ID NO. 39. In an embodiment, the siRNA telomerase upregulating polynucleotides of the present invention comprise a 3* deoxythymidine dinucleotide (dTdT) overhang to increase nuclease resistance.

[0056] In an embodiment, the one or more telomerase upregulating polynucleotides of the present invention comprise a first siRNA and a second siRNA wherein the nucleotide sequence of the first siRNA is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to the nucleotide sequence of SEQ ID NO. 26 and the nucleotide sequence of the second siRNA is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to the nucleotide sequence of SEQ ID NO. 229.

[0057] SEQ ID NO. 17 (siasTERCl): GCCUACGCCCUUCUCAGUUAG

[0058] SEQ ID NO. 18 (siasTERC2): GGCCGAGGCUUUUCCGCCCGC

[0059] SEQ ID NO. 19 (siasTERC3): AGAGCCCAACUCUUCGCGGUG

[0060] SEQ ID NO. 20 (siasTERC4): GCAGCUGACAUUUUUUGUUUG

[0061] SEQ ID NO. 21 (siasTERC5): CCGCCGCAGGUCCCCGGGAGG

[0062] SEQ ID NO. 22 (siasTERC6): AUCGGCGUUCCCCCCACCAAC

[0063] SEQ ID NO. 23 (siasTERC7): GCCUUCACGUCUCCUGCCAAU

[0064] SEQ ID NO. 24 (siasTERC8): GGUUCACAAGCCCCCAUUGCC

[0065] SEQ ID NO. 25 (siasTERC9): AUUUUGGCCGACUUUGGAGGU

[0066] SEQ ID NO. 26 (siasTERC10): CUGCCCAUUCAUUUUGGCCGA

[0067] SEQ ID NO. 27 (siasTERC11): UUAGCUUAUUUUCUUAAAGGA

[0068] SEQ ID NO. 28 (siasTERC12): GUUAAAUGAUCUCUCAAAAAA

[0069] SEQ ID NO. 29 (siasTERC13): UCAUUUAGAUCUUCUAAUUAA

[0070] SEQ ID NO. 30 (siasTERC14): AAGGUCUAAUUUUUCAAAAAA

[0071] SEQ ID NO. 31 (siasTERC15): UCUUUUCCGUCUUUCAUUAUG

[0072] SEQ ID NO. 32 (MG-si1): GAUGAUGUCUUAGUCAUCACAGUGU

[0073] SEQ ID NO. 33 (MG-si2): GGAAAAUUUGUUCCAACAAAAGUUA

[0074] SEQ ID NO. 34 (MG-si3): GAGGCCAACAUCUGGUCACAUCCCG

[0075] SEQ ID NO. 35 (E666-si 1): CCAGCAGGUACUUAUCAUGAAUCCU

[0076] SEQ ID NO. 36 (E666-si2): GUACACUGCGUUUCUGUGCACCAUC

[0077] SEQ ID NO. 37 (E666-si3): CCUGUUCUCUGCAAUCCUUCCAGUU

[0078] SEQ ID NO. 38 (E666-si4-1): GGCACCAGAGUGCAUUUACGA

[0079] SEQ ID NO. 39 (E666-si4-2): UUCUCUGAAGCAGCAGCAAUA

[0080] In an embodiment, the one or more telomerase upregulating polynucleotides of the present invention comprise a shRNA. In an embodiment, each of the one or more shRNA telomerase upregulating polynucleotides of the present invention comprises a first nucleotide sequence and a second nucleotide sequence wherein the second nucleotide sequence is reverse complementary to the first nucleotide sequence. In an embodiment, the nucleotide sequence of the first nucleotide sequence of each of the one or more telomerase upregulating shRNAs of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 19, SEQ ID NO. 20, SEQ ID NO. 21, SEQ ID NO. 22, SEQ ID NO. 23, SEQ ID NO. 24, SEQ ID NO. 25, SEQ ID NO. 26, SEQ ID NO. 27, SEQ ID NO. 28, SEQ ID NO. 29, SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 32, SEQ ID NO. 33, SEQ ID NO. 34, SEQ ID NO. 35, SEQ ID NO. 36, SEQ ID NO. 37, SEQ ID NO. 38, or SEQ ID NO. 39. In an embodiment, the shRNA of the present invention further comprises a loop connecting the first nucleotide sequence and the second nucleotide sequence. In an embodiment, the loop of the telomerase upregulating shRNA of the present invention comprises nucleotide sequence of about 2-9 bp such as about 2, 3, 4, 5, 6, 7, 8, or 9 bp. In an embodiment, nucleotide sequence of the loop of the telomerase upregulating shRNA of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 40, SEQ ID NO. 41, SEQ ID NO. 42, SEQ ID NO. 43, or SEQ ID NO. 44. In an embodiment, the loop may be cleaved off in RNA interference pathway.

[0081] SEQ ID NO. 40: UUGC

[0082] SEQ ID NO. 41 : CCUGACCCA

[0083] SEQ ID NO. 42: AAGCACA

[0084] SEQ ID NO. 43: UGUGCUU

[0085] SEQ ID NO. 44: UUG

[0086] In an embodiment, the one or more telomerase upregulating polynucleotides of the present invention comprise a first shRNA and a second shRNA wherein the nucleotide sequence of the first nucleotide sequence of the first shRNA is al least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 45 and the nucleotide sequence of the first nucleotide sequence of the second shRNA is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 46.

[0087] SEQ ID NO. 45: TCGGCCAAAATGAATGGGCAGTTGCCTGCCCATTCATTTTGGCCGA

[0088] SEQ ID NO. 46: TTAATTAGAAGATCTAAATGAGCAATCATTTAGATCTTCTAATTAA

[0089] In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention further comprises N-acetylgalactosamine (GalNAc) wherein the one or more telomerase upregulating polynucleotides of the present invention is conjugated to the GalNAc. In an embodiment, any embodiments of the telomerase upregulating siRNAs of the present invention further comprises GalNAc wherein the one or more telomerase upregulating siRNAs of the present invention is conjugated to the GalNAc. In an embodiment, any embodiments of the telomerase upregulating shRNAs of the present invention further comprises GalNAc wherein the one or more telomerase upregulating shRNAs of the present invention is conjugated to the GalNAc.

[0090] In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention upregulates the telomerase activity of a subject by at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 500%, about 750% or about 1000%. In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention restores the telomerase activity of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of the normal telomerase activity of the subject when healthy. In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention restores thetelomerase activity of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal telomerase activity of an average healthy human being. In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention restores the telomerase activity of a subject to al least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal telomerase activity of an average healthy human being having the same biometrics of the subject such as age, sex, height, weight etc... or a combination thereof.

[0091] In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention upregulates the expression of TERT, TERC, or a combination thereof of a subject by at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 500%, about 750% or about 1000%. In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention restores the expression of TERT, TERC, or a combination of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of the normal expression level of TERT, TERC, or a combination of the subject when healthy. In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention restores the expression of the TERT, TERC, or a combination of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal expression level of TERT, TERC, or a combination of an average healthy human being. In an embodiment, any embodiment of the telomerase upregulating polynucleotides of the present invention restores the expression of TERT, TERC, or a combination of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal expression level of TERT, TERC, or a combination of an average healthy human beinghaving the same biometrics of the subject such as age, sex, height, weight etc... or a combination thereof.

[0092] The present invention also provides a pharmaceutical composition comprising a nanoparticle encapsulating any embodiment of the one or more telomerase upregulating polynucleotides of the present invention. In an embodiment, the nanoparticle of the present invention comprises chemical nanoparticles such as but not limited to lipid nanoparticle, polymer nanoparticle, lipid-polymer hybrid nanoparticle and biological nanoparticles such as but not limited to liposome, exosome, virus, or virus-like particle.

[0093] In an embodiment, the nanoparticle comprises an exosome. In an embodiment, the exosome encapsulates one or more fusion proteins and one or more cargo RNA. In an embodiment, the cargo RNA comprises a package RNA and a packaging domain wherein the package RNA comprises any embodiment of the one or more telomerase upregulating polynucleotides of the present invention and wherein the packaging domain is capable of binding to the packaging protein of the fusion protein. In an embodiment, the fusion protein comprises an exosome associated transmembrane protein fused to a packaging protein. In an embodiment, the exosome associated transmembrane protein comprises CD9, CD37, CD53, CD63, CD68, CD81, CD82, LAMP-1, LAMP-2A, LAMP-2B, LAMP-2C, lactadherin, or PTGFRN. In an embodiment, the packaging protein comprises U1 a protein. In an embodiment, the packaging protein comprises a RNA-binding protein capable of binding tot the packaging domain such that the cargo RNA binds to the fusion protein via the RNA- binding protein and the packaging domain.

[0094] In an embodiment, the nanoparticle comprises an exosome prepared from an exosome-based packaging and delivery system. In an embodiment, the system comprises a low immunogenic exosome-based packaging and delivery system. In an embodiment, the exosome-based RNA package and delivery system comprises an exosome producing cell, a cargo RNA plasmid and one or more fusion protein plasmids, wherein the cargo RNA plasmid encodes a package RNA comprising any embodiment of the one or more telomerase upregulating polynucleotides of the present invention and a packaging domain capable of binding to the packaging protein of the fusion protein encoded by the one or more fusion protein plasmids. In an embodiment, the fusion protein expressed by the exosome producing cell based on the fusion protein plasmid comprises an exosome associated transmembrane protein fused to a packaging protein. In an embodiment, the exosome associated transmembrane protein comprises CD9, CD37, CD53, CD63, CD68, CD81, CD82, LAMP-1, LAMP-2A, LAMP-2B, LAMP-2C, lactadherin, or PTGFRN. In an embodiment, thepackaging protein comprises an RNA-binding protein. In an embodiment, the packaging protein is endogenous to a subject. Various embodiments for the recombinant fusion protein comprising an exosome associated transmembrane protein and a packaging protein were disclosed in PCT application no. PCT / US2021 / 026892 filed 12 April, 2021, the contents of which are incorporated by reference herein in its entirety. In an embodiment, the packaging protein comprises U1 a protein.

[0095] In an embodiment the one or more fusion proteins comprise CD63-Ula protein. In an embodiment, the nucleotide sequence of the CD63-U1 a fusion protein encoding plasmid of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 47. In an embodiment, the amino acid sequence of the CD63-Ula fusion protein is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 48. In an embodiment the one or more fusion proteins comprise CD81-U1 a protein. In an embodiment, the nucleotide sequence of the CD81-Ula fusion protein encoding plasmid of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 49. In an embodiment, the amino the acid sequence of the CD81-U la fusion protein is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 50. In an embodiment, the one or more fusion proteins comprise PTGFRN-Ula protein. In an embodiment, the nucleotide sequence of the PTGFRN-Ula fusion protein encoding plasmid of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 51. In an embodiment, the amino the acid sequence of the PTGFRN-Ula fusion protein is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 52.

[0096] SEQ ID NO. 47 (nt CD63-Ula):ATGGCGGTGGAAGGAGGAATGAAATGTGTGAAGTTCTTGCTCTACGTCCTCCTGCTGGCCTHTGCGCCTGTGCAGTGGGACTGATTGCCGTGGGTGTCGGGGCACAGCTTGTCCTGAGTCAGACCATAATCCAGGGGGCTACCCCTGGCTCTCTGTTGCCAGTGGTCATCATCGCAGTGGGTGTCTTCCTCTTCCTGGTGGCTTTTGTGGGCTGCTGCGGGGCCTGCAAGGAGAACTATTGTCTTATGATCACGTTTGCCATCnTCTGTCTCTTATCATGTTGGTGGAGGTGGCCGCAGCCATTGCTGGCTATGTGTTTAGAGATAAGGTGATGTCAGAGTTTAATAACAACTTCCGGCAGCAGATGGAGAATTACCCGAAAAACAACCACACTGCTTCGATCCTGGACAGGATGCAGGCAGATnTAAGTGCTGTGGGGCTGCTAACTACACAGATTGGGAGAAAATCCCTTCCATGTCGAAGAACCGAGTCCCCGACTCCTGCTGCATTAATGTTACTGTGGGCTGTGGGATTAATTTCAACGAGAAGGCGATCCATAAGGAGGGCTGTGTGGAGAAGATTGGGGGCTGGCTGAGGAAAAATGTGCTGGTGGTAGCTGCAGCAGCCCTTGGAATTGCTTTTGTCGAGGTTTTGGGAATTGTCTTTGCCTGCTGCCTCGTGAAGAGTATCAGAAGTGGCTACGAGGTGATGgaattcggcggaggcgggtccATGGCAGTTCCCGAGACCCGCCCTAACCACACTATTTATATCAACAACCTCAATGAGAAGATCAAGAAGGATGAGCTAAAAAAGTCCCTGTACGCCATCTTCTCCCAGTTTGGCCAGATCCTGGATATCCTGGTATCACGGAGCCTGAAGATGAGGGGCCAGGCCTTTGTCATCTTCAAGGAGGTCAGCAGCGCCACCAACGCCCTGCGCTCCATGCAGGGTTTCCCTTTCTATGACAAACCTATGCGTATCCAGTATGCCAAGACCGACTCAGATATCATTGCCAAGATGAAA

[0097] Uppercase sequence denotes CD63, lower case is linker sequence, and bold sequence is the U1 small nuclear ribonucleoprotein polypeptide A (U1 snRNP A).

[0098] SEQ ID NO. 48 (aa CD63-Ula):MAVEGGMKCVKFLLYVLLLAFCACAVGLIAVGVGAQLVLSQTnQGATPGSLLPWIIAVGVFLFLVAFVGCCGACKENYCLMITFAIFLSLIMLVEVAAAIAGYVFRDKVMSEFNNNFRQQMENYPKNNHTASILDRMQADFKCCGAANYTDWEKIPSMSKNRVPDSCCINVTVGCG1NFNEKA1HKEGCVEK1GGWLRKNVLVVAAAALG1AFVEVLG1VFACCLVKSIRSGYEVMefggggsMAVPETRPNHTIYINNLNEKIKKDELKKSLYAIFSQFGQILDILVSRSLKMRGQAFVIFKEVSSATNALRSMQGFPFYDKPMRIQYAKTDSDIIAKMK

[0099] Uppercase sequence denotes CD63, lower case is linker sequence, and bold sequence is the U1 small nuclear ribonucleoprotein polypeptide A (U1 snRNP A).[000100] SEQ ID NO. 49 (nt CD81-Ula):[000101] ATGTCCGGACTCAGATCTCGAGCTCAAGCTTCCGGAGTGGAGGGCTGCACCAAGTGCATCAAGTACCTGCTCTTCGTCTTCAATTTCGTCTTCTGGCTGGCTGGAGGCGTGATCCTGGGTGTGGCCCTGTGGCTCCGCCATGACCCGCAGACCACCAACCTCCTGTATCTGGAGCTGGGAGACAAGCCCGCGCCCAACACCTTCTATGTAGGCATCTACATCCTCATCGCTGTGGGCGCTGTCATGATGTTCGTTGGCTTCCTGGGCTGCTACGGGGCCATCCAGGAATCCCAGTGCCTGCTGGGGACGTTCTTCACCTGCCTGGTCATCCTGTTTGCCTGTGAGGTGGCCGCCGGCATCTGGGGCTTTGTCAACAAGGACCAGATCGCCAAGGATGTGAAGCAGTTCTATGACCAGGCCCTACAGCAGGCCGTGGTGGATGATGACGCCAACAACGCCAAGGCTGTGGTGAAGACCTTCCACGAGACGCTTGACTGCTGTGGCTCCAGCACACTGACTGCTTTGACCACCTCAGTGCTCAAGAACAATTTGTGTCCCTCGGGCAGCAACATCATCAGCAACCTCTTCAAGGAGGACTGCCACCAGAAGATCGATGACCTCTTCTCCGGGAAGCTGTACCTCATCGGCATTGCTGCCATCGTGGTCGCTGTGATCATGATCTTCGAGATGATCCTGAGCATGGTGCTGTGCTGTGGCATCCGGAACAGCTCCGTGTACgaattcggcggaggcgggtccATGGCAGTTCCCGAGACCCGCCCTAACCACACTATTTATATCAACAACCTCAATGAGAAGATCAAGAAGGATGAGCTAAAAAAGTCCCTGTACGCCATCTTCTCCCAGTTTGGCCAGATCCTGGATATCCTGGTATCACGGAGCCTGAAGATGAGGGGCCAGGCCTTTGTCATCTTCAAGGAGGTCAGCAGCGCCACCAACGCCCTGCGCTCCATGCAGGGTTTCCCTTTCTATGACAAACCTATGCGTATCCAGTATGCCAAGACCGACTCAGATATCATTGCCAAGATGAAATAA[000102] Uppercase sequence denotes CD81, lower case is linker sequence, and bold sequence is the U1 small nuclear ribonucleoprotein polypeptide A (U1 snRNP A).[000103] SEQ ID NO. 50 (aa CD81-Ula):[000104] MSGLRSRAQASGVEGCTKCIKYLLFVFNFVFWLAGGVILGVALWLRHDPQTTNLLYLELGDKPAPNTFYVGIYILIAVGAVMMFVGFLGCYGAIQESQCLLGTFFTCLVILFACEVAAGIWGFVNKDQIAKDVKQFYDQALQQAVVDDDANNAKAVVKTFHETLDCCGSSTLTALTTSVLKNNLCPSGSNnSNLFKEDCHQKIDDLFSGKLYLIGIAAIVVAVIM1FEM1LSMVLCCGIRNSSVYEFGGGGSMAVPETRPNHTIY1NNLNEKIKKDELKKSLYAIFSQFGQILDILVSRSLKMRGQAFVIFKEVSSATNALRSMQGFPFYDKPMRIQYAKTDSDIIAKMK[000105] SEQ ID NO. 51 (nt PTGFRN-Ula):[000106] ATGGGGCGCCTGGCCTCCAGGCCGCTGCTGCTGGCGCTCCTGTCGTTGGCTCTTTGCCGAGGGCGTGTGGTGAGAGTCCCCACAGCGACCCTGGTTCGAGTGGTGGGCACTGAGCTGGTCATCCCCTGCAACGTCAGTGACTATGATGGCCCCAGCGAGCAAAACTITGACTGGAGCTTCTCATCTTTGGGGAGCAGCTTTGTGGAGCTTGCAAGCACCTGGGAGGTGGGGTTCCCAGCCCAGCTGTACCAGGAGCGGCTGCAGAGGGGCGAGATCCTGTTAAGGCGGACTGCCAACGACGCCGTGGAGCTCCACATAAAGAACGTCCAGCCTTCAGACCAAGGCCACTACAAATGTTCAACCCCCAGCACAGATGCCACTGTCCAGGGAAACTATGAGGACACAGTGCAGGTTAAAGTGCTGGCCGACTCCCTGCACGTGGGCCCCAGCGCGCGGCCCCCGCCGAGCCTGAGCCTGCGGGAGGGGGAGCCCTTCGAGCTGCGCTGCACCGCCGCCTCCGCCTCGCCGCTGCACACGCACCTGGCGCTGCTGTGGGAGGTGCACCGCGGCCCGGCCAGGCGGAGCGTCCTCGCCCTGACCCACGAGGGCAGGTTCCACCCGGGCCTGGGGTACGAGCAGCGCTACCACAGTGGGGACGTGCGCCTCGACACCGTGGGCAGCGACGCCTACCGCCTCTCAGTGTCCCGGGCTCTGTCTGCCGACCAGGGCTCCTACAGGTGTATCGTCAGCGAGTGGATCGCCGAGCAGGGCAACTGGCAGGAAATCCAAGAAAAGGCCGTGGAAGTTGCCACCGTGGTGATCCAGCCATCAGTTCTGCGAGCAGCTGTGCCCAAGAATGTGTCTGTGGCTGAAGGAAAGGAACTGGACCTGACCTGTAACATCACAACAGACCGAGCCGATGACGTCCGGCCCGAGGTGACGTGGTCCTTCAGCAGGATGCCTGACAGCACCCTACCTGGCTCCCGCGTGTTGGCGCGGCTTGACCGTGATTCCCTGGTGCACAGCTCGCCTCATGTTGCTTTGAGTCATGTGGATGCACGCTCCTACCATTTACTGGTTCGGGATGTTAGCAAAGAAAACTCTGGCTACTATTACTGCCACGTGTCCCTGTGGGCACCCGGACACAACAGGAGCTGGCACAAAGTGGCAGAGGCCGTGTCTTCCCCAGCTGGTGTGGGTGTGACCTGGCTAGAACCAGACTACCAGGTGTACCTGAATGCTTCCAAGGTCCCCGGGTTTGCGGATGACCCCACAGAGCTGGCATGCCGGGTGGTGGACACGAAGAGTGGGGAGGCGAATGTCCGATTCACGGTTTCGTGGTACTACAGGATGAACCGGCGCAGCGACAATGTGGTGACCAGCGAGCTGCTTGCAGTCATGGACGGGGACTGGACGCTAAAATATGGAGAGAGGAGCAAGCAGCGGGCCCAGGATGGAGACTTTATTTTTrCTAAGGAACATACAGACACGTTCAATTTCCGGATCCAAAGGACTACAGAGGAAGACAGAGGCAATTATTACTGTGTTGTGTCTGCCTGGACCAAACAGCGGAACAACAGCTGGGTGAAAAGCAAGGATGTCTTCTCCAAGCCTGTTAACATA mTGGGCATTAGAAGATTCCGTGCTTGTGGTGAAGGCGAGGCAGCCAAAGCCTTTCTTTGCTGCCGGAAATACATTTGAGATGACTTGCAAAGTATCTTCCAAGAATATTAAGTCGCCACGCTACTCTGTTCTCATCATGGCTGAGAAGCCTGTCGGCGACCTCTCCAGTCCCAATGAAACGAAGTACATCATCTCTCTGGACCAGGATTCTGTGGTGAAGCTGGAGAATTGGACAGATGCATCACGGGTGGATGGCGTTGTTTTAGAAAAAGTGCAGGAGGATGAGTTCCGCTATCGAATGTACCAGACTCAGGTCTCAGACGCAGGGCTGTACCGCTGCATGGTGACAGCCTGGTCTCCTGTCAGGGGCAGCCHTGGCGAGAAGCAGCAACCAGTCTCTCCAATCCTATTGAGATAGACTTCCAAACCTCAGGTCCTATATTTAATGCTTCTGTGCATTCAGACACACCATCAGTAATTCGGGGAGATCTGATCAAATTGTTCTGTATCATCACTGTCGAGGGAGCAGCACTGGATCCAGATGACATGGCCTTTGATGTGTCCTGGTTTGCGGTGCACTCTTTTGGCCTGGACAAGGCTCCTGTGCTCCTGTCTTCCCTGGATCGGAAGGGCATCGTGACCACCTCCCGGAGGGACTGGAAGAGCGACCTCAGCCTGGAGCGCGTGAGTGTGCTGGAATTCTTGCTGCAAGTGCATGGCTCCGAGGACCAGGACTTTGGCAACTACTACTGTTCCGTGACTCCATGGGTGAAGTCACCAACAGGTTCCTGGCAGAAGGAGGCAGAGATCCACTCCAAGCCCGTnTTATAACTGTGAAGATGGATGTGCTGAACGCCTTCAAGTATCCCTTGCTGATCGGCGTCGGTCTGTCCACGGTCATCGGGCTCCTGTCCTGTCTCATCGGGTACTGCAGCTCCCACTGGTGTTGTAAGAAGGAGGTTCAGGAGACACGGCGCGAGCGCCGCAGGCTCATGTCGATGGAGATGGACgaattcggcggaggcgggtccATGGCAGTTCCCGAGACCCGCCCTAACCACACTATTTATATCAACAACCTCAATGAGAAGATCAAGAAGGATGAGCTAAAAAAGTCCCTGTACGCCATCTTCTCCCAGTTTGGCCAGATCCTGGATATCCTGGTATCACGGAGCCTGAAGATGAGGGGCCAGGCCTTTGTCATCTTCAAGGAGGTCAGCAGCGCCACCAACGCCCTGCGCTCCATGCAGGGTTTCCCTTTCTATGACAAACCTATGCGTATCCAGTATGCCAAGACCGACTCAGATATCATTGCCAAGATGAAATAG[000107] Uppercase sequence denotes PTGFRN, lower case is linker sequence, and bold sequence is the U1 small nuclear ribonucleoprotein polypeptide A (U1 snRNP A). [000108] SEQ ID NO. 52 (aa PRGFRN-Ula): [000109] MGRLASRPLLLALLSLALCRGRWRVPTATLVRWGTELVIPCNVSDYDGPSEQNFDWSFSSLGSSFVELASTWEVGFPAQLYQERLQRGEILLRRTANDAVELHI KNVQPSDQGHYKCSTPSTDATVQGNYEDTVQVKVLADSLHVGPSARPPPSLSLREG EPFELRCTAASASPLHTHLALLWEVHRGPARRSVLALTHEGRFHPGLGYEQRYHSGD VRLDTVGSDAYRLSVSRALSADQGSYRCIVSEWIAEQGNWQEIQEKAVEVATVVIQP SVLRAAVPKNVSVAEGKELDLTCNITTDRADDVRPEVTWSFSRMPDSTLPGSRVLARLDRDSLVHSSPHVALSHVDARSYHLLVRDVSKENSGYYYCHVSLWAPGHNRSWHKVAEAVSSPAGVGVTWLEPDYQVYLNASKVPGFADDPTELACRWDTKSGEANVRF TVSWYYRMNRRSDNWTSELLAVMDGDWTLKYGERSKQRAQDGDFIFSKEHTDTF NFRIQRTTEEDRGNYYCWSAWTKQRNNSWVKSKDVFSKPVNIFWALEDSVLVVK ARQPKPFFAAGNTFEMTCKVSSKNIKSPRYSVLIMAEKPVGDLSSPNETKYIISLDQDS VVKLENWTDASRVDGWLEKVQEDEFRYRMYQTQVSDAGLYRCMVTAWSPVRGS LWREAATSLSNPEIDFQTSGPIFNASVHSDTPSVIRGDLIKLFCnTVEGAALDPDDMA FDVSWFAVHSFGLDKAPVLLSSLDRKGIVTTSRRDWKSDLSLERVSVLEFLLQVHGS EDQDFGNYYCSVTPWVKSPTGSWQKEAEIHSKPVFITVKMDVLNAFKYPLLIGVGLS TVIGLLSCLIGYCSSHWCCKKEVQETRRERRRLMSMEMDEFGGGGSMAVPETRPNH TIYINNLNEKIKKDELKKSLYAIFSQFGQILDILVSRSLKMRGQAFVIFKEVSSATNALR SMQGFPFYDKPMRIQYAKTDSDIIAKMK[000110] In an embodiment, the cargo RNA comprises a package RNA comprising any embodiment of the one or more telomerase upregulating polynucleotides of the present invention and a packaging domain capable of binding to the packaging protein of the fusion protein. In an embodiment, the packaging domain comprises UR domain or L2 domain. In an embodiment the nucleotide sequence of the UR domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 53 and the nucleotidesequence of the L2 domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID No. 54.[000111] In an embodiment, the packaging domain further comprises stabilizing domains. In an embodiment, the stabilizing domain comprises OH domain and / or MorrisMotif domain wherein the nucleotide sequence of the OH domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 55 and the nucleotide sequence of the MorrisMotif domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 56.[000112] SEQ ID NO. 53 (UR): AATCCATTGCACTCCGGATT[000113] SEQ ID NO. 54 (L2): AATCCATTGCACTCCGGATTT[000114] SEQ ID NO. 55 (OH): CTGCAGATATCCAGCACAGTGGC[000115] SEQ ID NO. 56 (MorrisMotif): GCGCAGCGCGCGCAGCGC[000116] In an embodiment, the cargo RNA further comprises a SIRLOIN (SINE- derived nuclear RNA LOcalizatloN) nuclear localization sequences wherein the SIRLOIN is upstream of the UR or L2 sequence of the cargo RNA. In an embodiment, the nucleotide sequence of SIRLOIN is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 57.[000117] SEQ ID NO. 57 (SIRLOIN):CGCCTCCCGGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCGA[000118] In an embodiment, the exosome-based packaging and delivery system further comprises an argonaute 2 (Ago2)-encoding plasmid. In an embodiment, the Ago2 comprises S387 A mutation. Overexpression of Ago2 or the S387A mutant thereof increases the packaging efficiency of the cargo RNA into the exosomes in an exosome-producing cell. [000119] In an embodiment, the pharmaceutical composition comprising a nanoparticle encapsulating any embodiment of the one or more telomerase upregulating polynucleotides of the present invention further comprises a nanoparticle payload release enhancer. In an embodiment, the nanoparticle release enhancer enhances the release of payload from the nanoparticle by at least about 2-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to the same composition without release enhancer. In an embodiment, the nanoparticle payload release enhancer comprises a modified myoferlin protein. In an embodiment, the modified myoferlin protein comprises C2F, C2G, transmembrane domain, or a combination thereof. In an embodiment, the modified myoferlin protein consists of C2F, C2G, transmembrane domain, or a combination thereof. In an embodiment, the amino acid sequence of the C2F domain is atleast about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 58. In an embodiment, the amino acid sequence of the C2G domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 59. In an embodiment, the amino acid sequence of the transmembrane domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 60.[000120] SEQ ID NO. 58 (C2F):QFRELPDSVPQECTVRIYIYRGLELQPQDNNGLCDPYIKHWKKVIEDRDHYIPNTLNPVFGRMYELSCYLPQEKDLKISVYDYDTFTRDEKVGETIIDLENRFLSRFGSHCGIPEE YCVSGV[000121] SEQ ID NO. 59 (C2G):PFNTTPRKAKKYYLRVIIWNTKDVILDEKSITGEEMSDIYVKGWVPGNEENKQKTDVHYRSLDGEGNFNWRFVFPFDYLPAEQLCIVAKKEHFWSIDQTEFRIPPRLIIQIWDNDKFSLDDYLGFLELDLRH[000122] SEQ ID NO. 60 (transmembrane domain):PDLKAMNPLKAKTASLFEQKSMKGWWPCYAEKDGARVMAGKVEMTLE1LNEKEADERPAGKGRDEPNMNPKLDLPNRPETSFLWFTNPCKTMKHVWRRFKWVIIGLLFLLILLLFVAV[000123] In an embodiment, the modified myoferlin protein comprises C2A, FerA, FerB, DysFN, transmembrane domain, or a combination thereof. In an embodiment, the modified myoferlin protein consists of C2A, FerA, FerB, DysFN, transmembrane domain. In an embodiment, the amino acid sequence of the C2A domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 61. In an embodiment, the amino acid sequence of the FerA domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 62. In an embodiment, the amino acid sequence of the FerB is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 63. In an embodiment, the DysFN domain comprises DysFN- 1, DysFN-2, or a combination thereof. In an embodiment, the amino acid sequence of the DysFN- 1 is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 64. In an embodiment, the amino acid sequence of the DysFN-2 is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 65. In an embodiment, the amino acid sequence of the transmembrane domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 60.[000124] SEQ ID NO. 61 (C2A): MLRVIVESASNIPKTKFGKPDPIVSVIFKDEKKKTKKVDNELNPVWNEILEFDLRGIPLDFSSSLGIIVKDFETIGQNKLIGTATVALKDLTGDQSRSLPYKLISLLNERGQDTGATIDLVIGYDPPSAPHPNDLS[000125] SEQ ID NO. 62 (FerA):LQTNIEALKSGIQGKIPANQLAELWLKLIDEVIEDTRYTLPLTEGKANVTVLDTQIRK[000126] SEQ ID NO. 63 (FerB):[000127] WLDKLMQLTEEPQNSMPDIIIWMIRGEKRLAYARIPAHQVLYSTSGENASGKYCGKTQTIFLKYPQEKNNGP[000128] SEQ ID NO. 64 (DysFN-1):AVEKKFNSFAEGTFTVFAEMYENQALMFGKWGTSGLVGRHKFSDVTGKIKLKREFFLP[000129] SEQ ID NO. 65 (DysFN-2): DPERSLLTEADAGHTEFTDEVYQNESRYPGGDWKPAEDTYTDANGDKAASPSELTC P[000130] In an embodiment, any embodiment of the modified myoferlin protein of the present invention further comprises C2B, C2C, C2D, C2E domains, or a combination thereof. In an embodiment, the amino acid sequence of the C2B domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 66. In an embodiment, the amino acid sequence of the C2C domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 67. In an embodiment, the amino acid sequence of the C2D is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 68. In an embodiment, the amino acid sequence of the C2E is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 69.[000131] SEQ ID NO. 66 (C2B): PQDFQIRVRVIEGRQLSGNNIRPVVKVHVCGQTHRTRIKRGNNPFFDELFFYNVNMT PSELMDEIISIRVYNSHSLRADCLMGEFKIDVGFVYDEPGHAVMRKWLLLNDP[000132] SEQ ID NO. 67 (C2C):TFLLKIYRAEDIPQMDDAFSQTVKEIFGGNADKKNLVDPFVEVSFAGKKVCTNIIEKNANPEWNQVVNLQIKFPSVCEKIKLTIYDWDRLTKNDVVGTTYLHLSKIAASGGEVEDFSSSGTGAASYTVNTGETEVGFVPTFGPCYLNLYGSPREYTGFPDPYDE[000133] SEQ ID NO. 68 (C2D):TPIVSCNFDRVYIYHLRCYVYQARNLLALDKDSFSDPYAHICFLHRSKTTEIIHSTLNPTWDQTIIFDEVEIYGEPQTVLQNPPKVIMELFDNDQVGKDEFLGRSIFSPVVKLNSEMDITPKLLWHPVMNGDKA[000134] SEQ ID NO. 69 (C2E):RNMKNFQMASITSPSLVVECGGERVESVVIKNLKKTPNFPSSVLFMKVFLPKEELYM PPLVIKVIDHRQFGRKPVVG[000135] In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2A domain wherein the amino acid sequence of the C2A domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 61. In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2B domain wherein the amino acid sequence of the C2B domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ DI NO. 66. In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2C domain wherein the amino acid sequence of the C2C domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ DI NO. 67. In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2D domain wherein the amino acid sequence of the C2D domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 68. In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2E domain wherein the amino acid sequence of the C2E domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 69. In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2F domain wherein the amino acid sequence of the C2F domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 58. In an embodiment, the modified myoferlin protein of the present invention does not comprise the C2G domain wherein the amino acid sequence of the C2G domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 59. In an embodiment, the modified myoferlin protein of the present invention does not comprise the FerA domain wherein the amino acid sequence of the FerA domain is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 62. In an embodiment, the modified myoferlin protein of the present invention does not comprise the FerB domain wherein the amino acid sequence of the FerB domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 63. In an embodiment, the modified myoferlin protein of the present invention does not comprise the DysFN-1 domain wherein the amino acid sequence of the DysFN-1domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 64. In an embodiment, the modified myoferlin protein of the present invention does not comprise the DysFN-2 domain wherein the amino acid sequence of the DysFN-2 domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 65. In an embodiment, the modified myoferlin protein of the present invention does not comprise the transmembrane domain wherein the amino acid sequence of the transmembrane domain is at least about 80%, about 85%, about 90%, about 95% or about 100%identical or similar to SEQ ID NO. 60.[000136] In an embodiment, the amino acid sequence of the modified myoferlin protein of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 70.[000137] SEQ ID NO. 70 (C2F-C2G):MVPAPPRQFRELPDSVPQECTVRIYIVRGLELQPQDNNGLCDPYIKITLGKKVIEDRD HYIPNTLNPVFGRMYELSCYLPQEKDLKISVYDYDTFTRDEKVGETnDLENRFLSRFG SHCGIPEEYCVSGVNTWRDQLRPTQLLQNVARFKGFPQPILSEDGSRIRYGGRDYSLD EFEANKILHQHLGAPEERLALH1LRTQGLVPEHVETRTLHSTFQPNISQGKLQMWVD VFPKSLGPPGPPFNITPRKAKKYYLRVnWNTKDVILDEKSITGEEMSDIYVKGWVPGNEENKQKTDVHYRSLDGEGNFNWRFVFPFDYLPAEQLCIVAKKEHFWSIDQTEFRIPPRLIIQIWDNDKFSLDDYLGFLELDLRHTIIPAKSPEKCRLDMIPDLKAMNPLKAKTASLFEQKSMKGWWPCYAEKDGARVMAGKVEMTLEILNEKEADERPAGKGRDEPNMNPKLDLPNRPETSFLWFTNPCKTMKFIVWRRFKWVnGLLFLLILLLFVAVLLYSLPNYLSMKIVKPNVYPYDVPDYA[000138] The present invention also provides a modified myoferlin protein encoding- polynucleotide encoding any embodiment of the modified myoferlin protein of the present invention. In an embodiment, the modified myoferlin protein encoding polynucleotide encodes a C2A domain, a C2B domain, a C2C domain, a C2D domain, a C2E domain, a C2F domain, a C2G domain, a FerA domain, a FerB domain, a DysFN-1 domain, a DysFN-2 domain, a transmembrane domain, or a combination thereof. In an embodiment, the C2A domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 71. In an embodiment, the C2B domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 72. In an embodiment, the C2C domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at leastabout 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 73. In an embodiment, the C2D domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 74. In an embodiment, the C2E domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 75. In an embodiment, the C2F domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 76. In an embodiment, the C2G domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 77. In an embodiment, the FerA domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 78. In an embodiment, the FerB domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 79. In an embodiment, the DysFN-1 domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 80. In an embodiment, the DysFN-2 domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 81. In an embodiment, the transmembrane domain is encoded by a polynucleotide wherein nucleotide sequence of the polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 82. [000139] SEQ ID NO. 71 (nt C2A): ATGCTGCGAGTGATTGTGGAATCTGCCAGCAATATCCCTAAAACGAAATTTGGCAAGCCGGATCCTATTGTTTCTGTCATmTAAGGATGAGAAAAAGAAAACAAAGAAAGTTGATAATGAATTGAACCCTGTCTGGAATGAGATTTTGGAGTTTGACTTGAGGGGTATACCACTGGACTTTTCATCTTCCCTTGGGATTATTGTGAAAGATTTTGAGACAATTGGACAAAATAAATTAATTGGCACGGCGACTGTAGCCCTGAAGGACCTGACTGGTGACCAGAGCAGATCCCTGCCGTACAAGCTGATCTCCCTGCTAAATGAAAGAGGGCAAGATACTGGGGCCACCATTGACTTGGTGATCGGCTATGATCCGCCTTCTGCTCCACATCCAAATGACCTGAGC[000140] SEQ ID NO. 72 (nt C2B):CCACAGGACTTCCAGATCCGCGTCCGAGTGATTGAGGGCCGACAGTTAAGTGGCAACAACATAAGGCCTGTGGTCAAAGTTCACGTCTGTGGCCAGACACACCGAACAAGAATCAAGAGAGGAAACAACCCTTTTTrTGATGAGTTGTTTTTCTACAATGTCAACATGACCCCTTCTGAATTGATGGATGAGATCATCAGCATCCGGGTTTATAATTCTCACTCTCTGCGGGCAGATTGTCTGATGGGGGAATTTAAGATTGATGTTGGATTTGTTTATGATGAACCTGGCCATGCTGTCATGAGAAAGTGGCTTCTTCTCAATGACCCG[000141] SEQ ID NO. 73 (ntC2C):ACCTTCTTGCTGAAAATCTACCGAGCTGAGGACATCCCCCAGATGGATGATGCCTTCTCACAGACAGTAAAGGAAATATTTGGAGGCAATGCAGATAAGAAAAATCTCGTGGATCCTTTTGTAGAAGTTTCCTTTGCTGGAAAAAAGGTTTGTACAAACATAATTGAGAAGAATGCAAACCCAGAGTGGAATCAGGTCGTCAATCTTCAGATCAAGTTTCCTTCAGTGTGTGAAAAAATAAAACTAACAATATATGACTGGGACCGTCTTACTAAAAATGATGTAGTTGGAACAACATATCTACACCTCTCTAAAATTGCTGCCTCTGGTGGGGAAGTGGAAGATTTCTCATCTTCGGGAACTGGGGCTGCATCATATACAGTAAACACAGGAGAAACAGAGGTAGGCTTTGTTCCAACGTTTGGACCTTGTTACCTGAATCTTTATGGAAGCCCCAGAGAGTACACGGGATTCCCAGACCCCTATGATGAG[000142] SEQ ID NO. 74 (ntC2D):ACCCCCATTGTTTCCTGCAATTTTGACAGAGTCTACATCTACCATCTGCGCTGCTATGTCTATCAAGCCAGAAACCTCTTGGCTTTAGATAAGGATAGCTTTTCAGATCCATATGCTCATATCTGTTTCCTCCATCGGAGCAAAACCACTGAGATCATCCATTCAACCCTGAATCCCACGTGGGACCAAACAATTATATTCGATGAAGTTGAAATCTATGGGGAACCCCAAACAGTTCTACAGAATCCACCCAAAGTTATCATGGAACTTTTTGACAATGACCAAGTGGGCAAAGATGAATTTTTAGGACGAAGCATTTTCTCTCCTGTGGTGAAACTGAACTCAGAAATGGACATCACACCCAAACTTCTCTGGCACCCAGTAATGAATGGAGACAAAGCC[000143] SEQ ID NO. 75 (ntC2E):AGAAATATGAAAAACTTCCAGATGGCTTCTATCACATCCCCCAGTCTTGTTGTGGAGTGTGGAGGAGAAAGGGTGGAATCGGTGGTGATCAAAAACCTTAAGAAGACACCCAACTTTCCAAGTTCTGTTCTCTTCATGAAAGTGTTCTTGCCCAAGGAGGAATTGTACATGCCCCCACTGGTGATCAAGGTCATCGACCACAGGCAGTTTGGGCGGAAGCCTGTCGTCGGC[000144] SEQ ID NO. 76 (ntC2F):CAGTTTCGGGAATTACCTGACAGCGTCCCACAGGAATGCACGGTTAGGATTTACATTGTTCGAGGCTTAGAGCTCCAGCCCCAGGACAACAATGGCCTGTGTGACCCTTACATAAAAATAACACTGGGCAAAAAAGTCATTGAAGACCGAGATCACTACATTCCCAACACTCTCAACCCAGTCTTTGGCAGGATGTACGAACTGAGCTGCTACTTACCTCAAGAAAAAGACCTGAAAATTTCTGTCTATGATTATGACACCTTTACCCGGGATGAAAAAGTAGGAGAGACAATTATTGATCTGGAAAACCGATTCCTTTCCCGCTTTGGGTCCCACTGCGGCATACCAGAGGAGTACTGTGTTTCTGGAGTC[000145] SEQ ID NO. 77 (nt C2G):CCTTTCAACATCACACCCCGGAAAGCCAAGAAATACTACCTGCGTGTGATCATCTGGAACACCAAGGATGTTATCTTGGATGAGAAAAGCATCACAGGAGAGGAAATGAGTGACATCTACGTCAAAGGCTGGGTTCCTGGCAATGAAGAAAACAAACAGAAAACAGATGTCCATTACAGATCTTTGGATGGTGAAGGGAATnTAACTGGCGATTTGTTTTCCCGTTTGACTACCTTCCAGCCGAACAACTCTGTATCGTTGCGAAAAAAGAGCATTTCTGGAGTATTGACCAAACGGAATTTCGAATCCCACCCAGGCTGATCATTCAGATATGGGACAATGACAAGTTTTCTCTGGATGACTACTTGGGTTTCCTAGAACTTGACTTGCGTCAC[000146] SEQ ID NO. 78 (nt FerA):CTGCAAACAAATATAGAAGCTCTAAAATCAGGGATACAAGGTAAAATTCCTGCAAACCAGCTGGCTGAATTGTGGCTGAAGCTGATAGATGAAGTTATAGAAGACACGAGATACACGTTGCCTCTCACAGAAGGAAAAGCCAACGTCACAGTTCTCGATACTCAGATCCGAAAG[000147] SEQ ID NO. 79 (nt FerB):TGGCTTGATAAATTAATGCAGCTGACTGAAGAGCCACAGAACAGCATGCCTGACATCATCATCTGGATGATCCGGGGAGAGAAGAGACTGGCCTATGCACGAATTCCCGCACATCAGGTCTTGTACTCCACCAGTGGTGAGAATGCATCTGGAAAATACTGTGGGAAAACCCAAACCATCTTTCTGAAGTATCCACAGGAGAAAAACAACGGGCCA[000148] SEQ ID NO. 80 (nt DysFN-1):GCTGTGGAGAAGAAGTTTAACAGCTTCGCAGAAGGAACTTTCACCGTCnTGCTGAAATGTATGAAAATCAAGCTCTCATGTTTGGAAAATGGGGTACTTCTGGATTAGTAGGACGTCATAAGTTTTCTGATGTCACAGGAAAAATAAAACTCAAGAGGGAATTTITTCTGCCT[000149] SEQ ID NO. 81 (nt DysFN-2):GATCCTGAAAGAAGCTTGCTGACTGAGGCAGATGCAGGTCACACGGAGTTCACTGATGAAGTCTACCAGAACGAGAGCCGCTACCCCGGGGGCGACTGGAAGCCGGCCGAGGACACCTACACGGATGCGAACGGCGATAAAGCAGCATCACCCAGCGAGTTGACTTGTCCT[000150] SEQ ID NO. 82 (nt TM): CCGGACCTCAAAGCCATGAACCCCCTTAAAGCCAAGACAGCCTCCCTCTTTGAGCAGAAGTCCATGAAAGGATGGTGGCCATGCTACGCAGAGAAAGATGGCGCCCGCGTAATGGCTGGGAAAGTGGAGATGACATTGGAAATCCTCAACGAGAAGGAGGCCGACGAGAGGCCAGCCGGGAAGGGGCGGGACGAACCCAACATGAACCCCAAGCTGGACTTACCAAATCGACCAGAAACCTCCTTCCTCTGGTTCACCAACCCATGCAAGACCATGAAGTTCATCGTGTGGCGCCGCTTTAAGTGGGTCATCATCGGCTTGCTGTTCCTGCTTATCCTGCTGCTCTTCGTGGCCGTG[000151] In an embodiment, the modified myoferlin protein of the present invention further comprises a connexin 43 protein, wherein said dual protein comprising the modified myoferlin protein and the connexin 43 protein can be used to greatly enhance the efficacy and delivery of the RNA or nucleic acid payloads. In an embodiment, the connexin 43 protein comprises a S368A mutation. In an embodiment, amino acid sequence of the connexin 43 protein of the present invention is at least about 80%, about 85%, about 90%, about 95% or about 100% identical or similar to SEQ ID NO. 83.[000152] SEQ ID NO. 83 (connexin 43):MATTMGDWSALGKLLDKVQAYSTAGGKVWLSVLFIFRILLLGTAVESAWGDEQSA FRCNTQQPGCENVCYDKSFPISHVRFWVLQIIFVSVPTLLYLAHVFYVMRKEEKLNK KEEELKVAQTDGVNVDMHLKQIEIKKFKYGIEEHGKVKMRGGLLRTYnSILFKSIFE VAFLLIQWYIYGFSLSAVYTCKRDPCPHQVDCFLSRPTEKTIFnFMLVVSLVSLALNII ELFYVFFKGVKDRVKGKSDPYHATSGALSPAKDCGSQKYAYFNGCSSPTAPLSPMSP PGYKLVTGDRNNSSCRNYNKQASEQNWANYSAEQNRMGQAGSTISNSHAQPFDFP DDNQNSKKLAAGHELQPLAIVDQRPSSRAASRASSRPRPDDLEI[000153] The present invention further provides a dual protein polynucleotide encoding the dual protein comprising a modified myoferlin protein of the present invention fused to a connexin 43 protein S368A mutant wherein nucleotide sequence of the dual protein polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 84.[000154] SEQ ID NO. 84:ATGGTGCCAGCCCCTCCCAGACAGTTTCGGGAATTACCTGACAGCGTCCCACAGGAATGCACGGTTAGGATTTACATTGTTCGAGGCTTAGAGCTCCAGCCCCAGGACAACAATGGCCTGTGTGACCCTTACATAAAAATAACACTGGGCAAAAAAGTCATTGAAGACCGAGATCACTACATTCCCAACACTCTCAACCCAGTCTTTGGCAGGATGTACGAACTGAGCTGCTACTTACCTCAAGAAAAAGACCTGAAAATTTCTGTCTATGATTATGACACCTTTACCCGGGATGAAAAAGTAGGAGAGACAATTATTGATCTGGAAAACCGATTCCTTTCCCGCTTTGGGTCCCACTGCGGCATACCAGAGGAGTACTGTGTTTCTGGAGTCAATACCTGGCGAGATCAACTGAGACCAACACAGCTGCTTCAAAATGTCGCCAGATTCAAAGGCTTCCCACAACCCATCCTTTCCGAAGATGGGAGTAGAATCAGATATGGAGGACGAGACTACAGCTTGGATGAATTTGAAGCCAACAAAATCCTGCACCAGCACCTCGGGGCCCCTGAAGAGCGGCTTGCTCTTCACATCCTCAGGACTCAGGGGCTGGTCCCTGAGCACGTGGAAACAAGGACTTTGCACAGCACCTTCCAGCCCAACATTTCCCAGGGAAAACTTCAGATGTGGGTGGATGTTTTCCCCAAGAGTTTGGGGCCACCAGGCCCTCCTTTCAACATCACACCCCGGAAAGCCAAGAAATACTACCTGCGTGTGATCATCTGGAACACCAAGGATGTTATCTTGGATGAGAAAAGCATCACAGGAGAGGAAATGAGTGACATCTACGTCAAAGGCTGGGTTCCTGGCAATGAAGAAAACAAACAGAAAACAGATGTCCATTACAGATCTTTGGATGGTGAAGGGAATTTTAACTGGCGATTTGTTTTCCCGTTTGACTACCTTCCAGCCGAACAACTCTGTATCGTTGCGAAAAAAGAGCATTTCTGGAGTATTGACCAAACGGAATTTCGAATCCCACCCAGGCTGATCATTCAGATATGGGACAATGACAAGTTTTCTCTGGATGACTACTTGGGTTTCCTAGAACTTGACTTGCGTCACACGATCATTCCTGCAAAATCACCAGAGAAATGCAGGTTGGACATGATTCCGGACCTCAAAGCCATGAACCCCCTTAAAGCCAAGACAGCCTCCCTCTTTGAGCAGAAGTCCATGAAAGGATGGTGGCCATGCTACGCAGAGAAAGATGGCGCCCGCGTAATGGCTGGGAAAGTGGAGATGACATTGGAAATCCTCAACGAGAAGGAGGCCGACGAGAGGCCAGCCGGGAAGGGGCGGGACGAACCCAACATGAACCCCAAGCTGGACTTACCAAATCGACCAGAAACCTCCTTCCTCTGGTTCACCAACCCATGCAAGACCATGAAGTTCATCGTGTGGCGCCGCTTTAAGTGGGTCATCATCGGCTTGCTGTTCCTGCTTATCCTGCTGCTCTTCGTGGCCGTGCTCCTCTACTCnTGCCGAACTATTTGTCAATGAAGATTGTAAAGCCAAATGTGTACCCATACGACGTCCCAGACTACGCTTAGGCCCCTCTCCCTCCCCCCCCCCTAACGTTACTGGCCGAAGCCGCTTGGAATAAGGCCGGTGTGCGTTTGTCTATATGTTATnTCCACCATATTGCCGTCTnTGGCAATGTGAGGGCCCGGAAACCTGGCCCTGTCTTCTTGACGAGCATrCCTAGGGGTCTTrCCCCTCTCGCCAAAGGAATGCAAGGTCTGTTGAATGTCGTGAAGGAAGCAGTTCCTCTGGAAGCTrCTTGAAGACAAACAACGTCTGTAGCGACCCTTTGCAGGCAGCGGAACCCCCCACCTGGCGACAGGTGCCTCTGCGGCCAAAAGCCACGTGTATAAGATACACCTGCAAAGGCGGCACAACCCCAGTGCCACGTTGTGAGTTGGATAGTTGTGGAAAGAGTCAAATGGCTCTCCTCAAGCGTATTCAACAAGGGGCTGAAGGATGCCCAGAAGGTACCCCATTGTATGGGATCTGATCTGGGGCCTCGGTGCACATGCTTTACATGTGTTTAGTCGAGGTTAAAAAAACGTCTAGGCCCCCCGAACCACGGGGACGTGGTTTTCCTTTGAAAAACACGATGATAATATGGCCACAACCATGGGTGACTGGAGCGCCTTAGGCAAACTCCTTGACAAGGTTCAAGCCTACTCAACTGCTGGAGGGAAGGTGTGGCTGTCAGTACTTTTCATTTTCCGAATCCTGCTGCTGGGGACAGCGGTTGAGTCAGCCTGGGGAGATGAGCAGTCTGCCTTTCGTTGTAACACrCAGCAACCTGGTTGTGAAAATGTCTGCTATGACAAGTCTTTCCCAATCTCTCATGTGCGCTTCTGGGTCCTGCAGATCATATTTGTGTCTGTACCCACACTCTTGTACCTGGCTCATGTGTTCTATGTGATGCGAAAGGAAGAGAAACTGAACAAGAAAGAGGAAGAACTCAAGGTTGCCCAAACTGATGGTGTCAATGTGGACATGCACTTGAAGCAGATTGAGATAAAGAAGTTCAAGTACGGTATTGAAGAGCATGGTAAGGTGAAAATGCGAGGGGGGTTGCTGCGAACCTACATCATCAGTATCCTCTTCAAGTCTATCTTTGAGGTGGCCTTCTrGCTGATCCAGTGGTACATCTATGGATTCAGCTTGAGTGCTGTTTACACTTGCAAAAGAGATCCCTGCCCACATCAGGTGGACTGTTTCCTCTCTCGCCCCACGGAGAAAACCATCTTCATCATCTTCATGCTGGTGGTGTCC7FTGGTGTCCCTGGCCTTGAATATCATTGAACTCTTCTATGTTTTCTTCAAGGGCGTTAAGGATCGGGTTAAGGGAAAGAGCGACCCTTACCATGCGACCAGTGGTGCGCTGAGCCCTGCCAAAGACTGTGGGTCTCAAAAATATGCTTATTTCAATGGCTGCTCCTCACCAACCGCTCCCCTCTCGCCTATGTCTCCTCCTGGGTACAAGCTGGTTACTGGCGACAGAAACAATTCTTCTTGCCGCAATTACAACAAGCAAGCAAGTGAGCAAAACTGGGCTAATTACAGTGCAGAACAAAATCGAATGGGGCAGGCGGGAAGCACCATCTCTAACTCCCATGCACAGCCTHTGATTTCCCCGATGATAACCAGAATTCAAAAAAACTAGCTGCTGGACATGAATTACAGCCACTAGCCATTGTGGACCAGCGACCTTCAAGCAGAGCCGCCAGTCGTGCCAGCAGCAGACCTCGGCCTGATGACCTGGAGATCTGA[000155] In an embodiment, the various components comprising the fusion protein or any variant thereof of the present invention, the Ago2 protein or any variant thereof of the present invention, the modified myoferlin protein or any variant thereof of the present invention, or the connexin 43 protein or any variant thereof of the present invention that could be incorporated or encapsulated to the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention for the enhanced delivery and uptake of said nanoparticle in a subject can be incorporated or encapsulated to the nanoparticle in all possible combinations. In an embodiment, the nanoparticle encapsulatingthe one or more telomerase upregulating polynucleotides of the present invention further comprises a fusion protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention further comprises an Ago2 protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention further comprises a modified myoferlin protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention further comprises a connexin 43 protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention does not comprise a fusion protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention does not comprise an Ago2 protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention does not comprise a modified myoferlin protein or any variant thereof of the present invention. In an embodiment, the nanoparticle encapsulating the one or more telomerase upregulating polynucleotides of the present invention does not comprise a connexin 43 protein or any variant thereof of the present invention. In an embodiment where the nanoparticle comprises an exosome encapsulating one or more telomerase upregulating polynucleotides expression of the present invention and a fusion protein comprising an exosome-associated transmembrane protein fused to a packaging protein of the present invention, the one or more telomerase upregulating polynucleotides is fused to a packaging domain or any variant thereof of the present invention. In an embodiment, the packaging domain or any variant thereof of the present invention comprises UR, L2, the OH domain, the MorrisMotif domain, the nuclear localization sequence SIRLOIN, or a combination thereof.[000156] In an embodiment, any embodiment of the nanoparticle encapsulating the telomerase upregulating polynucleotides of the present invention upregulates the telomerase activity of a subject by at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 500%, about 750% or about 1000%. In an embodiment, any embodiment of the nanoparticle encapsulating the telomerase upregulatingpolynucleotides of the present invention restores the telomerase activity of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of the normal telomerase activity of the subject when healthy. In an embodiment, any embodiment of the nanoparticle encapsulating the telomerase upregulating polynucleotides of the present invention restores the telomerase activity of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal telomerase activity of an average healthy human being. In an embodiment, any embodiment of the nanoparticle encapsulating the telomerase upregulating polynucleotides of the present invention restores the telomerase activity of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal telomerase activity of an average healthy human being having the same biometrics of the subject such as age, sex, height, weight etc... or a combination thereof. [000157] In an embodiment, any embodiment of the nanoparticle encapsulating any embodiment of the telomerase upregulating polynucleotides of the present invention upregulates the expression of TERT, TERC, or a combination thereof of a subject by at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 500%, about 750% or about 1000%. In an embodiment, any embodiment of the nanoparticle encapsulating any embodiment of the telomerase upregulating polynucleotides of the present invention restores the expression of TERT, TERC, or a combination thereof of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of the normal expression level of TERT, TERC, or a combination thereof of the subject when healthy. In an embodiment, any embodiment of the nanoparticle encapsulating any embodiment of the telomerase upregulating polynucleotides of the present invention restores the expression of TERT, TERC, or a combination thereof of a subject to at least about 1 %, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%,about 90% or about 100% of normal expression level of TERT, TERC, or a combination thereof of an average healthy human being. In an embodiment, any embodiment of the nanoparticle encapsulating any embodiment of the telomerase upregulating polynucleotides of the present invention restores the expression of TERT, TERC, or a combination thereof of a subject to at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100% of normal expression level of TERT, TERC, or a combination thereof of an average healthy human being having the same biometrics of the subject such as age, sex, height, weight etc... or a combination thereof. [000158] The present invention provides a method of increasing the expression ofTERT, TERC, or a combination thereof in a subject comprising the administration of a therapeutically effective amount of any embodiment of the one or more telomerase upregulating polynucleotides of the present invention to the subject In an embodiment, the subject is diagnosed with telomerase-associated disease. In an embodiment, the telomerase- associated disease comprises telomeropathies, aging-associated disease, cellular senescence- associated disease, or a combination thereof. In an embodiment, the telomerase-associated disease comprises TERT- or TERC-associated chronic lung disease such as but not limited to chronic obstructive pulmonary disease, lung cancer, or idiopathic pulmonary fibrosis, TERT- or TERC-associated chronic liver disease such as but not limited to liver fibrosis or cirrhosis, or TERT- or TERC-associated bone marrow failure (BMF) such as but not limited to dyskeratosis congenita (DC), or aplastic anemia.[000159] The present invention also provides a method of treatment of a telomerase- associated disease of a subject comprising the step of increasing the telomerase expression of the subject. In an embodiment, the step of increasing the telomerase expression of a subject of the present invention comprises increasing the expression of TERT, TERC, or a combination of a subject. In an embodiment, the step of increasing the expression of TERT, TERC, or a combination comprises the administration of a therapeutically effective amount of any embodiment of the one or more telomerase upregulating polynucleotides of the present invention to the subject. In an embodiment, the step of increasing telomerase expression comprises the administration of any embodiment of the nanoparticle encapsulating one or more telomerase upregulating polynucleotides expression of the present invention to the subject[000160] In an embodiment, the telomerase-associated disease comprises telomeropathies, aging-associated disease, cellular senescence-associated disease, or acombination thereof. In an embodiment, the telomerase-associated disease comprises TERT- or TERC-associated chronic lung disease such as but not limited to chronic obstructive pulmonary disease, lung cancer, or idiopathic pulmonary fibrosis, TERT- or TERC- associated chronic liver disease such as but not limited to liver fibrosis or cirrhosis, or TERT- or TERC-associated bone marrow failure (BMP) such as but not limited to dyskeratosis congenita (DC), or aplastic anemia.[000161] In an embodiment, the method of treatment of a telomerase-associated disease in a subject comprises the step of administering a therapeutic effective amount of any embodiment of the pharmaceutical composition comprising one or more telomerase upregulating polynucleotides expression of the present invention to the subject. In an embodiment, the method of treating a telomerase-associated disease comprises the step of administering a therapeutic effective amount of any embodiment of the pharmaceutical composition comprising any embodiment of the nanoparticle encapsulating one or more telomerase upregulating polynucleotides expression of the present invention to the subject. The pharmaceutical compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic, vaginal, rectal, intranasal, transdermal), oral, or parenteral. Parenteral administration includes intravenous drip, subcutaneous, intraperitoneal or intramuscular injection, or intrathecal or intraventricular administration. The route and site of administration may be chosen to enhance delivery or targeting of the disrupting agent comprising a site-specific targeting moiety to a particular location. For example, to target liver cells, intravenous injection may be used.[000162] The pharmaceutical compositions may be administered in the form of any embodiment of the exosomes of the present invention. As used herein the term “exosome” refers to a cell-derived small (between 20-300 nm in diameter, more preferably 40-200 nm in diameter) vesicle comprising a membrane that encloses an internal space, and which is generated from said cell by direct plasma membrane budding or by fusion of the late endosome with the plasma membrane. The exosome comprises lipid or fatty add and polypeptide and further comprises the inhibitory nucleic adds described herein as a payload. The exosome can be derived from a producer cell, and isolated from the producer cell based on its size, density, biochemical parameters, or a combination thereof. Exosomes can be directly loaded with exogenous nucldc acids or drugs by electroporation, lipofection, sonication and contact with caldum chloride. Alternatively, purified exosomes may be loaded ex vivo by, for example, electroporation.[000163] Exosomes of the present invention can be produced from a cell grown in vitro or a body fluid of a subject. When exosomes are produced from in vitro cell culture, various producer cells, e.g., HEK293 cells, Chinese hamster ovary (CHO) cells, or mesenchymal stem cells (MSCs), can be used.[000164] Hie pharmaceutical compositions may also be formulated by incorporation of the inhibitory nucleic acids described herein into adenoviruses or adeno-associated viruses (AAVs), formulated with cell-penetrating peptides, lentiviral vectors, polymers, dendrimers, or prepared as siRNA bioconjugates such as the GalNAc-siRNA conjugate delivery platform. [000165] In an embodiment, if using the exosomes or a vector as a vehicle to deliver siRNA, the candidate siRNAs are delivered as shRNAs. Both siRNAs and shRNAs can target and repress viruses and are functionally equivalent. When the candidate siRNAs are delivered as shRNAs they are derived from a cell system and packaged into exosomes or a vector (AAV or Lentiviral vector) as described above.[000166] In an embodiment, an shRNA may be provided in an expression cassette containing a promoter contiguously linked to an siRNA as described herein. In embodiments, the promoter is a pollI or a pollll promoter, such as a U6 promoter (e.g., a mouse U6 promoter) or a Hl promoter. In embodiments, the expression cassette further contains a marker gene. In embodiments, the promoter is a poll! promoter. In embodiments, the promoter is a tissue-specific promoter. In embodiments, the promoter is an inducible promoter. In embodiments, the promoter is a pollll promoter. In embodiments, the promoter is U6 or Hl promoter.[000167] Also provided is a vector containing an expression cassette described herein. Examples of appropriate vectors include adenoviral, lentiviral, adeno-associated viral (AAV), poliovirus, herpes simplex virus (HSV), or murine Maloney-based viral vectors. In an embodiment, the vector is an adeno-associated virus (AAV) vector.[000168] Inan embodiment, a shRNA molecule comprises paired RNA sequences and a loop portion positioned between the paired RNA sequences so as to form the hairpin. The loop can vary in length. In some embodiments the loop is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides in length. In certain embodiments, the loop is 18 nucleotides in length. The hairpin structure can also contain 3 ' and / or 5 " overhang portions. In some embodiments, the overhang is a 3 " and / or a 5 " overhang 0, 1, 2, 3, 4 or 5 nucleotides in length. The nucleotide sequence of the loop region may vary and could be, forexample, (5’-GCAA-3’), (5’-GCGC-3’) or (5’-TTGC-3’) or other sequences as will be well understood by the skilled person.[000169] Hie pharmaceutical compositions described herein may be administered in dosages sufficient to inhibit the expression of the target gene or the biological activity of nontranslated target sequences (e.g. regulatory sequences) in a cell, tissue or organism under treatment. The specific dosages of the inhibitory nucleic acids described herein administered to a given subject will depend on factors such as the route of administration and physical characteristics of the subject (including health status) and so forth. For example, the appropriate dosage of a given pharmaceutical composition comprising the inhibitory nucleic acids described herein may depend on a variety of factors including, but not limited to, a subject's physical characteristics (e.g. age, weight, sex), the progression (i.e. pathological state) of a given coronavirus infection, and other factors that will be readily recognised by one skilled in the art. Various general considerations that may be considered when determining an appropriate dosage are described, for example, in Gennaro et al. (Eds), (1990), “Remington's Pharmaceutical Sciences”, Mack Publishing Co., Easton, Pennsylvania, USA; and Gilman et al. (Eds), (1990), “Goodman And Gilman’s: The Pharmacological Bases of Therapeutics”, Pergamon Press. Non-limiting examples of suitable dosages of the inhibitory nucleic acids described herein include those in the range of 0.01 to 200 milligrams per kilogram body weight of the recipient per day, 1 to 50 mg / kg body weight per day, 1 to 40 mg / kg body weight per day, 1 to 30 mg / kg body weight per day, 1 to 30 mg / kg body weight per day, 1 to 10 mg / kg body weight per day, 1 to 5 mg / kg body weight per day, 1 to 3 mg / kg body weight per day, 1 to 2 mg / kg body weight per day, 0.1 to 1 mg / kg body weight per day, 0.1 to 0.9 mg / kg body weight per day, 0.1 to 0.8 mg / kg body weight per day, 0.1 to 0.7 mg / kg body weight per day, 0.1 to 0.6 mg / kg body weight per day, 0.1 to 0.5 mg / kg body weight per day, 0.1 to 0.4 mg / kg body weight per day, 0.1 to 0.3 mg / kg body weight per day, 0.1 to 0.2 mg / kg body weight per day, 0.01 to 0.1 mg / kg body weight per day, 0.01 to 0.05 mg / kg body weight per day, 0.01 to 0.02 mg / kg body weight per day, and 0.005 to 0.01 mg / kg body weight per day.[000170] Those of ordinary skill in the art will be able, by routine experimentation, to determine an effective, non-toxic amount of the pharmaceutical compositions and / or inhibitory nucleic acids described herein to include in a dosage or in a series of dosages to achieve the desired therapeutic outcome.[000171] Typically, in therapeutic applications, the treatment would be for the duration of the infection, disease state or condition. Further, it will be apparent to one of ordinary skillin the art that the optimal quantity and spacing of individual dosages will be determined by the nature and extent of the infection, disease state or condition being treated, the form, route and site of administration, and the nature of the particular individual being treated. Such optimum conditions can also be determined using conventional techniques.[000172] In many instances, it will be desirable to have several or multiple administrations of a pharmaceutical composition described herein. For example, they may be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more times. The administrations may be from about one to about twelve week intervals, and in certain embodiments from about one to about four week intervals. Periodic re-administration may be desirable in the case of recurrent exposure to a particular pathogen targeted by a pharmaceutical composition described herein. [000173] It will also be apparent to one of ordinary skill in the art that the optimal course of treatment can be ascertained using conventional course of treatment determination tests.[000174] Suitable techniques for introduction of the inhibitory nucleic acids described herein into cells, tissues, and organisms include various carrier systems, vectors and reagents. Non-limiting examples include lipid nanoparticles (LNP), micelles, nucleic-acid-lipid particles, lipoplexes, liposomes, nucleic acid polymers, single chemical entity conjugates, virosomes, virus like particles (VLP), and mixtures thereof.[000175] Pharmaceutical compositions of the present invention may be administered in any suitable way, such as, for example, intravenously, buccally, parenterally, intranasally, orally, sublingually, or topically. Accordingly, the administration may be topical, pulmonary (e.g. by inhalation or insufflation of aerosols or powders including with a nebulizer), intranasal, intratracheal, epidermal, transdermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial (e.g. intraparenchymal, intrathecal or intraventricular) administration. [000176] In an embodiment, the pharmaceutical composition is adapted for intranasal administration.[000177] In an embodiment, a pharmaceutical composition of the present invention is formulated as a direct-acting nasal spray. In an embodiment, a nasal spray can be selfadministered at point-of-care.[000178] It is to be understood that both the foregoing general description and detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. In general, the terms used in the disclosure should not be construed to limit the technology to the specific embodiments disclosed in the specification, unless the abovedetailed description explicitly defines such terms. Accordingly, the actual scope of the technology encompasses the disclosed embodiments and all equivalent ways of practicing or implementing the technology.[000179] EXAMPLES[000180] Materials and methods[000181] The siRNAs were transfected into cells at lOOnM final concentration using Lipofectamine 2000 (L2K). Cell RNAs were collected at 72 or 96 hrs post-transfection using Qiagen RNAeasy kit with DNAse treatment The resultant RNAs were converted to cDNA using Thermo fisher Reverse Transcription kit. The cDNAs were then assessed for differential gene expression relative to beta actin using the primers from Table 1 by quantitative PCR.[000182] Table 1 - PCT primers used for RNA detection[000183] Results[000184] Example 1 - TERC-activating siRNAs[000185] Towards the goal of activating the TERT / TERC pathway, we surveyed theTERC and TERT genomic locus and observed several antisense long non-coding RNAs (IncRNA) which have been annotated and might be endogenous regulators of TERC (Figure 1) or TERT expression (Figure 2). Antisense IncRNAs have been well reported in the literature to control gene expression by a myriad of different mechanisms, including blocking miRNA targeting to protein coding genes and targeting epigenetic regulatory proteins to particular loci in the genome. We therefore surmised that targeting the TERC and TERT associated IncRNAs using siRNAs could be a viable means to de-repressing TERC and TERT expression, essentially activating TERC / TERT. To determine the ability to first activate TERC by repressing the various IncRNAs associated with the TERC locus (Figure 1), we developed several siRNAs (Table 2) targeted to these putative regulatory IncRNAs(Figure 3). The siRNAs are targeted to TERC associated antisense RNAs as a means to “derepress” and activate TERC expression. HEK293 cells were screened with siRNAs and the effects of these siRNAs on TERC expression determined by qRTPCR (Figure 4). Notably, we found siRNAs (2, 10 and 13) resulted in increased TERC expression (Figures 4 & 5), with siasTERC2 targeting (EST CV370609), siasTERClO targeting (ESTs AW207347,AI825849, AW136367, AI380754, AW293800) and siasTERC13 targeting (ESTBM695101)(Figure 6).[000186] Table 2 siRNAs targeted to TERC associated antisense IncRNAs.[000187] Example 2 - TERT-activating siRNAs[000188] Next, we assessed the TERT locus and found candidate IncRNAs that were antisense to the TERT gene (Figure 2). To determine the ability to activate TERT by repressing the various IncRNAs associated with the TERT locus (Figure 2), we developed several siRNAs C targeted to these putative regulatory IncRNAs (Table 3, Figure 7). The siRNAs are targeted to TERT associated antisense RNAs as a means to “de-repress” andactivate TERT expression. HEK293 cells were screened with siRNAs and the effects of these siRNAs on TERT expression determined by qRTPCR (Figures 8-9). Notably, we found two siRNAs, E666-si14-land E666-sil4-2, both targeted to regulate the IncRNA(ENST00000666708.1) resulted in increased TERT expression (Figure 10). Collectively, these data demonstrate that siRNA targeting of TERC and TERT regulatory IncRNAs is one means to activating TERC / TERT expression.[000189] Example 3 - Exosome delivery of TERT / TERC-activating siRNAs[000190] More recently we generated stable HEK293 shRNA-exosome producing cells.We engineered HEK293 cells to produce shRNA exosomes (shEVs) targeted to activateTERC by co-transfecting the dual shRNA expressing vector with the RNex packaging system(Table 4, Figure 11). We next drug selected these transfected cells to generate a stable shRNA producing cell and assessed these cell generated exosomes using a transwell assay(Figure 12) for activation of TERC. Our system of plasmids including the packer p24 and enhancer p54 (Table 4) were functionally required in all formulations to ultimately enhance shRNA packaging and endosomal release, as determined by shRNA knockdown and activation of target genes in recipient cell lines (Figure 13). One stable cell lines containing dual expressed shRNAs shlO and shl3 targeted to TERC regulatory IncRNAs was generated and found to activate TERC expression in recipient cells (Figure 13), as determined by qRT-PCR using TERC specific primers (Table 1). These shRNAs were developed to contain a7bp hairpin loop, but can also be used with 4bp, 5bp, 6bp, 8bp, and 9bp loops. The shRNA10 targets AW207347, AI825849, AW136367, AI380754, AW293800 while shRNA 13 targets BM695101. These data demonstrate that shRNAs targeted to regulatory IncRNAs that control TERC expression can be packaged and delivered by exosomes to target recipient cells.[000191] Table 3 siRNAs targeted TERT associated antisense IncRNAs.[000192] Table 4 Plasmids required to engineer cell line into shRNA-EV producing system. Plasmids were co-transfected at equal molar ratios and drug selected to generate stable shRNA-EV producer cells.[000193] It can be appreciated by those skilled in the art that changes could be made to the examples described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular examples disclosed, but it is intended to cover modifications within the spirit and scope of the present invention as defined by the appended claims.

Claims

1. A pharmaceutical composition comprising one or more telomerase upregulating polynucleotides, wherein the nucleotide sequence of each of the one or more telomerase upregulating polynucleotides is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 26, SEQ ID NO. 29, SEQ ID NO. 19, SEQ ID NO. 20, SEQ ID NO. 21, SEQ ID NO. 22, SEQ ID NO. 23, SEQ ID NO.24, SEQ ID NO. 25, SEQ ID NO. 17, SEQ ID NO. 27, SEQ ID NO. 28, SEQ ID NO.18, SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 32, SEQ ID NO. 33, SEQ ID NO.34, SEQ ID NO. 35, SEQ ID NO. 36, SEQ ID NO. 37, SEQ ID NO. 38, SEQ ID NO.39, or a combination thereof.

2. The pharmaceutical composition of claim 1, wherein the one or more telomerase upregulating polynucleotides upregulate the expression level of TERT, TERC, or a combination thereof by regulating expression level of one or more antisense regulator of TERT, TERC, or a combination thereof.

3. The pharmaceutical composition of claim 2, wherein nucleotide sequence of the antisense RNA regulator of TERT, TERC, or a combination thereof is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to the nucleotide sequence of AW207347 (SEQ ID NO. 1), AI825849 (SEQ ID NO. 2), AW136367 (SEQ ID NO. 3), AI380754 (SEQ ID NO. 4), AW293800 (SEQ ID NO. 5), BM695101 (SEQ ID NO. 6), CV370609 (SEQ ID NO. 7), MG677549 (SEQ ID NO. 8), BF802688 (SEQ ID NO. 9), AA748707 (SEQ ID NO. 10), AA811084 (SEQ ID NO. 11), AI824948 (SEQ ID NO. 12), AW270031 (SEQ ID NO. 13), AW276315 (SEQ ID NO. 14), DB345416 (SEQ ID NO. 15), or ENST00000666708.1 (SEQ ID NO. 16).

4. The pharmaceutical composition of claim 1, wherein the one or more telomerase upregulating polynucleotides comprise small interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA), ribozyme, deoxyribozyme, aptamer, or a combination thereof.

5. The pharmaceutical composition of claim 1, wherein the one or more telomerase upregulating polynucleotides comprise one or more of a first telomerase upregulating polynucleotide and one or more of a second telomerase upregulating polynucleotide wherein nucleotide sequence of the one or more of the first telomerase upregulating polynucleotide is at least about 80%, about 85%, about 90%, about 95% or about 100% identical to the nucleotide sequence of SEQ ID NO. 26 and nucleotide sequence of the one or more of the second telomerase upregulating polynucleotide is at least about 80%,about 85%, about 90%, about 95% or about 100% identical to SEQ ID NO. 29.

6. The pharmaceutical composition of claim 1 further comprising a nanoparticle wherein the nanoparticle encapsulates the one or more telomerase upregulating polynucleotides.

7. The pharmaceutical composition of claim 6, wherein the nanoparticle comprises lipid nanoparticle, polymer nanoparticle, lipid-polymer hybrid nanoparticle, liposome, exosome, or virus or virus-like particle.

8. The pharmaceutical composition of claim 7, further comprising a fusion protein and a packaging domain wherein the nanoparticle comprises exosome, wherein the one or more telomerase upregulating polynucleotides is fused to the packaging domain, wherein the fusion protein comprises an exosome-associated transmembrane protein fused to a packaging protein, wherein the packaging domain comprises an UR domain, a L2 domain, or a combination thereof and wherein the packaging domain is capable of binding with the packaging protein.

9. The pharmaceutical composition of claim 8, wherein the one or more telomerase upregulating polynucleotides is further fused to a nuclear localization sequence SIRLOIN.

10. The pharmaceutical composition of claim 8, wherein the fusion protein comprises CD63-Ula, CD81-Ula, PTGFRN-Ula, or a combination thereof.

11. The pharmaceutical composition of claim 8, wherein the exosome further comprises an Ago2 protein or a S387A mutant thereof.

12. The pharmaceutical composition of claim 8, wherein the exosome further comprises a modified myoferlin protein.

13. The pharmaceutical composition of claim 12, wherein the modified myoferlin protein comprises C2F, C2G, transmembrane domain of the myoferlin protein, or a combination thereof.

14. The pharmaceutical composition of claim 12, wherein the modified myoferlin protein further comprises a connexin 43 protein or a S3 68A mutant thereof.

15. The pharmaceutical composition of claim 8, wherein the exosome is prepared using an exosome-based packaging and delivery system comprising one or more cargo RNA encoding plasmid and one or more fusion protein encoding plasmids wherein the one or more cargo RNA encoding plasmids each encodes the one or more telomerase upregulating polynucleotides fused to the packaging domain and wherein the one or more fusion protein encoding plasmids each encodes the fusion protein comprising an exosome-associated transmembrane protein fused to a packaging protein.

16. The pharmaceutical composition of claim 15, wherein the exosome-based delivery and packaging system further comprises a plasmid encoding a modified myoferlin protein, a connexin 43 protein or a S368A mutant thereof, or a combination thereof.

17. A method of increasing the expression of TERT, TERC, or a combination thereof in a subject comprising the step of administration of a therapeutically effective amount of the pharmaceutical composition of claim 1 to the subject.

18. The method of claim 17, wherein the subject suffers from a telomerase-associated disease comprising telomeropathies, aging-associated disease, cellular senescence-associated disease, or a combination thereof.

19. The method of claim 18, wherein the telomerase-associated disease comprises TERT-or TERC-associated chronic lung disease comprising chronic obstructive pulmonary disease, lung cancer, or idiopathic pulmonary fibrosis, TERT- or TERC-associated chronic liver disease such as but not limited to liver fibrosis or cirrhosis, or TERT- or TERC-associated bone marrow failure (BMF) such as but not limited to dyskeratosis congenita (DC), or aplastic anemia.

20. A method of treatment of telomerase-associated disease of a subject comprising the step of administration of a therapeutically effective amount of the pharmaceutical composition of claim 1 to the subject.

21. The method of claim 20, wherein the telomerase-associated disease comprises telomeropathies, aging-associated disease, cellular senescence-associated disease, or a combination thereof.

22. The method of claim 20, wherein the telomerase-associated disease comprises TERT-or TERC-associated chronic lung disease such as but not limited to chronic obstructive pulmonary disease, lung cancer, or idiopathic pulmonary fibrosis, TERT- or TERC-associated chronic liver disease such as but not limited to liver fibrosis or cirrhosis, or TERT- or TERC-associated bone marrow failure (BMF) such as but not limited to dyskeratosis congenita (DC), or aplastic anemia.

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