Compositions of modified TREM and uses thereof

Abstract

The present invention relates generally to tRNA-based effector molecules (TREMs) that contain an asialoglycoprotein receptor (ASGPR) binding moiety, and compositions and methods related thereto.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 130,373, U.S. Provisional Patent Application No. 63 / 130,374, U.S. Provisional Patent Application No. 63 / 130,375, U.S. Provisional Patent Application No. 63 / 130,377, U.S. Provisional Patent Application No. 63 / 130,381, and U.S. Provisional Patent Application No. 63 / 130,387, each filed on December 23, 2020. The entire contents of each of the foregoing applications are incorporated herein by reference. [Background technology]

[0002] tRNAs are complex RNA molecules that have several functions, including the ability to initiate and elongate proteins. Summary of the Invention [Means for solving the problem]

[0003] In particular, the present disclosure features tRNA-based effector molecule (TREM) entities that include an asialoglycoprotein receptor (ASGPR)-binding moiety, as well as compositions and methods of use thereof. The ASGPR-binding moiety can be conjugated to a nucleobase within the TREM entity, within an internucleotide linkage of the TREM entity, or at a terminus (e.g., the 5' or 3' end) of the TREM entity. In some embodiments, the TREM entity includes a TREM, a TREM core fragment, or a TREM fragment. In some embodiments, the nucleobase includes adenine, thymine, cytosine, guanosine, or uracil, or a mutant or modified form thereof.

[0004] In some aspects, a TREM entity (e.g., a TREM) described herein comprises a sequence of the formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2] (A), where, independently, the TREM comprises an ASGPR-binding moiety. In some embodiments, the ASGPR-binding moiety comprises an ASGPR carbohydrate and an ASGPR linker. In some embodiments, the ASGPR-binding moiety comprises a galactose (Gal) and / or N-acetylgalactosamine (GalNAc) moiety. In some embodiments, the ASGPR-binding moiety comprises multiple Gal and / or GalNAc moieties (e.g., 2, 3, 4, 5, 6, 7, 8, or more Gal and / or GalNAc moieties). In some embodiments, the ASGPR-binding moiety comprises a tri-antennary GalNAc moiety. In certain embodiments, the TREM further comprises a chemical modification (eg, a phosphothiorate internucleotide linkage or a 2'-modification on the ribose moiety within the TREM).

[0005] In some embodiments, the ASGPR-binding moiety is present on a nucleobase within a nucleotide in the TREM. In some embodiments, the ASGPR-binding moiety is present on the 5' end of the TREM. In some embodiments, the ASGPR-binding moiety is present on the 3' end of the TREM.

[0006] In some embodiments, the ASGPR binding moiety is present in a TREM domain selected from L1, ASt domain 1, L2, DH domain, L3, ACH domain, VL domain, TH domain, L4, and ASt domain 2. In some embodiments, the ASGPR binding moiety is present in the L1 region. In some embodiments, the ASGPR binding moiety is present in the AST domain 1. In some embodiments, the ASGPR binding moiety is present in the L2 region. In some embodiments, the ASGPR binding moiety is present in the DH domain. In some embodiments, the ASGPR binding moiety is present in the L3 region. In some embodiments, the ASGPR binding moiety is present in the ACH domain. In some embodiments, the ASGPR binding moiety is present in the VL domain. In some embodiments, the ASGPR binding moiety is present in the TH domain. In some embodiments, the ASGPR binding moiety is present in the L4 region. In some embodiments, the ASGPR binding moiety is present in AST domain 2.

[0007] In some embodiments, a TREM comprising an ASGPR-binding moiety retains the ability to assist in protein synthesis, be charged by synthetases, be bound by elongation factors, incorporate amino acids into peptide chains, assist in elongation, and / or assist initiation. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises at least X contiguous nucleotides that have no chemical modifications, where X is greater than 10. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises no more than 5, 10, or 15 nucleotides of a type (e.g., A, T, C, G, or U) that does not comprise a chemical modification. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, or 80 or fewer nucleotides of a type that does not comprise a chemical modification (e.g., A, T, C, G, or U). In some embodiments, a TREM comprising an ASGPR-binding moiety comprises at least X consecutive nucleotides that comprise a chemical modification, where X is greater than 10. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises 5, 10, or more than 15 nucleotides of a type that comprises a chemical modification (e.g., A, T, C, G, or U). In some embodiments, a TREM comprising an ASGPR-binding moiety comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, or 80 nucleotides of a type (e.g., A, T, C, G, or U) that comprises a chemical modification. In some embodiments, the chemical modification is a natural or non-natural chemical modification (e.g., a phosphothiolate internucleotide linkage or a 2'-modification on the ribose moiety in the TREM). In some embodiments, the chemical modification comprises a fluorophore.

[0008] In another aspect, the TREMs, or compositions thereof, comprising an ASGPR-binding moiety described herein may be used to regulate production parameters (e.g., expression parameters and / or signaling parameters) of an RNA corresponding to a nucleic acid sequence comprising an endogenous open reading frame (ORF) with a premature termination codon (PTC), or a polypeptide encoded by such a nucleic acid sequence.

[0009] In another aspect, the TREM comprising an ASGPR-binding moiety, or a composition thereof, described herein, can be used in a method for regulating a production parameter of an mRNA corresponding to an endogenous open reading frame (ORF) in a subject, or a polypeptide encoded by such an endogenous open reading frame (ORF), that contains a premature termination codon (PTC), comprising contacting the subject with a TREM comprising an ASGPR-binding moiety, or a composition thereof, in an amount and / or for a period of time sufficient to regulate the production parameter of the mRNA or polypeptide, thereby regulating the production parameter in the subject, wherein the TREM comprising an ASGPR-binding moiety has an anticodon that pairs with a codon having a first sequence. In certain embodiments, the production parameter comprises a signaling parameter and / or an expression parameter, e.g., as described herein.

[0010] In another aspect, the TREM comprising an ASGPR-binding moiety, or a composition thereof, described herein, can be used in a method of treating a subject having an endogenous open reading frame (ORF) containing a premature stop codon (PTC), the method comprising: providing a TREM comprising an ASGPR-binding moiety, or a composition thereof, wherein the TREM comprises an anticodon that pairs with the PTC in the ORF; and contacting the subject with the TREM comprising an ASGPR-binding moiety, or a composition thereof, in an amount and / or for a period of time sufficient to treat the subject, thereby treating the subject. In some embodiments, the PTC comprises UAA, UGA, or UAG.

[0011] In another aspect, the TREMs comprising an ASGPR-binding moiety, or compositions thereof, described herein, can be used in a method of treating a subject having a disease or disorder associated with a premature termination codon (PTC), the method comprising: providing a TREM or composition comprising an ASGPR-binding moiety described herein; contacting the subject with a TREM comprising an ASGPR-binding moiety, or composition thereof, in an amount and / or for a period of time sufficient to treat the subject, thereby treating the subject. In some embodiments, the PTC comprises UAA, UGA, or UAG. In some embodiments, the disease or disorder associated with a PTC is a disease or disorder described herein, such as cancer or a monogenic disorder.

[0012] Additional features of the aforementioned TREM entities (e.g., any of the TREMs, TREM core fragments, TREM fragments, TREM compositions, preparations, methods of making the TREM compositions and preparations, and methods of using the TREM compositions and preparations) include one or more of the listed features.

[0013] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein which equivalents are intended to be encompassed by the embodiments recited below. [Brief explanation of the drawings]

[0014] [Figure 1] 1 is an image showing ASGPR-expressing U2OS cells transfected with an exemplary TREM comprising an ASGPR-binding moiety described herein. In this experiment, uptake of a TREM comprising a SEQ ID NO: 650 backbone with an ASGPR-binding moiety at various positions along its sequence and conjugated to Cy3 was monitored and visualized by fluorescence microscopy. [Figure 2] 1A-1J are graphical representations of the fluorescence microscopy results of Figures 1A-1J, where the results are shown as the mean intensity of the concentration of oligo (nM) given to the cells. [Figure 3] 1 is an image showing ASGPR-expressing U2OS cells transfected with an exemplary TREM comprising an ASGPR-binding moiety described herein. In this experiment, uptake of a TREM comprising a SEQ ID NO: 650 backbone with an ASGPR-binding moiety at various positions along its sequence and conjugated to Cy3 was monitored and visualized by fluorescence microscopy. [Figure 4] 3A-3H are graphical representations of the fluorescence microscopy results. Results are shown as the mean intensity of the concentration of oligo (nM) given to the cells. [Figure 5] 1 is an image showing ASGPR-expressing U2OS cells transfected with an exemplary TREM comprising an ASGPR-binding moiety described herein. In this experiment, uptake of a TREM comprising a SEQ ID NO: 622 scaffold with an ASGPR-binding moiety at various positions along its sequence and conjugated to Cy3 was monitored and visualized by fluorescence microscopy. [Figure 6] 5A-5J are graphical representations of the fluorescence microscopy results of Figures 5A-5J, where the results are shown as the mean intensity of the concentration of oligo (nM) given to the cells. [Figure 7] 1 is an image showing the uptake of exemplary TREMs comprising the ASGPR-binding moieties described herein by primary human hepatocytes. In this experiment, the uptake of TREMs comprising SEQ ID NO: 650 backbone with ASGPR-binding moieties at various positions along the sequence and conjugated to Cy3 was monitored and visualized by fluorescence microscopy. [Figure 8] 7A-7J are graphical representations of the fluorescence microscopy results of Figures 7A-7J, where the results are shown as the mean intensity of the concentration of oligo (nM) given to the cells. [Figure 9] 1 shows an image showing the uptake of exemplary TREMs comprising the ASGPR binding moieties described herein by primary human hepatocytes. In this experiment, the uptake of TREMs comprising SEQ ID NO: 653 backbone with ASGPR binding moieties at various positions along the sequence and conjugated to Cy3 was monitored and visualized by fluorescence microscopy. [Figure 10]9A-9H are graphical representations of the fluorescence microscopy results of Figures 9A-9H, where the results are shown as the mean intensity of the concentration of oligo (nM) given to the cells. [Figure 11] 1 shows an image showing the uptake of an exemplary TREM comprising an ASGPR-binding moiety described herein by primary human hepatocytes. In this experiment, the uptake of a TREM comprising a SEQ ID NO: 622 scaffold with an ASGPR-binding moiety at various positions along its sequence and conjugated to Cy3 was monitored and visualized by fluorescence microscopy. [Figure 12] 11A-11J are graphical representations of the fluorescence microscopy results of Figures 11A-11J, where the results are shown as the mean intensity of the concentration of oligo (nM) given to the cells. [Figure 13] Graph showing the results of exemplary TREM uptake by ASGPR-expressing U2OS cells transfected with nLUC premature termination codon (PTC) reporter.Exemplary TREM containing SEQ ID NO: 650 backbone containing ASGPR-binding moieties positioned along the sequence was transfected using RNAiMAX transfection reagent.Results are shown as fold change relative to mock (no TREM) sample. [Figure 14] Graph showing the results of exemplary TREM uptake by ASGPR-expressing U2OS cells transfected with nLUC premature termination codon (PTC) reporter.Exemplary TREM containing SEQ ID NO: 653 backbone containing ASGPR-binding moiety positioned along the sequence was transfected using RNAiMAX transfection reagent.Results are shown as fold change relative to mock (no TREM) sample. [Figure 15] Graph showing the results of exemplary TREM uptake by ASGPR-expressing U2OS cells transfected with nLUC premature termination codon (PTC) reporter.Exemplary TREM containing SEQ ID NO: 622 backbone containing ASGPR-binding moieties positioned along the sequence was transfected using RNAiMAX transfection reagent.Results are shown as fold change relative to mock (no TREM) sample. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present disclosure features tRNA-based effector molecule (TREM) entities (e.g., TREMs, TREM core fragments, and TREM fragments) that contain an asialoglycoprotein receptor (ASGPR)-binding moiety, as well as compositions and related methods of use. As disclosed herein, TREM entities (e.g., TREMs) are complex molecules that can mediate various cellular processes. Pharmaceutical TREM compositions, e.g., TREMs that contain an ASGPR-binding moiety, can be administered to cells, tissues, or subjects to modulate these functions.

[0016] definition "Acceptor stem domain (AStD)," as the term is used herein, refers to a domain that binds to an amino acid. In one embodiment, an AStD comprises ASt domain 1 and ASt domain 2. For example, ASt domain 1 is at or near the 5' end of a TREM, and ASt domain 2 is at or near the 3' end of a TREM. An AStD comprises an RNA sequence sufficient to accept an amino acid, e.g., its cognate or non-cognate amino acid, and mediate the transfer of the amino acid (AA) in the initiation or elongation of a polypeptide chain, e.g., when present in an otherwise wild-type tRNA. Typically, an AStD comprises a 3'-terminal adenosine (CCA) for acceptor stem loading, which is part of synthetase recognition. In one embodiment, an AStD has at least 75, 80, 85, 85, 90, 95, or 100% identity to a naturally occurring AStD, e.g., an AStD encoded by a nucleic acid in Table 1. In some embodiments, a TREM may comprise a fragment or analog of an AStD, e.g., an AStD encoded by a nucleic acid in Table 1, where the fragment in some embodiments has AStD activity and in other embodiments does not have AStD activity. One of skill in the art can determine an appropriate corresponding sequence for any of the domains, stems, loops, or other sequence features described herein from a sequence encoded by a nucleic acid in Table 1. For example, one of skill in the art can determine a sequence corresponding to an AStD from a tRNA sequence encoded by a nucleic acid in Table 1. In some embodiments, the ASGPR-binding moiety is present within the AStD. In some embodiments, the ASGPR-binding moiety is bound to a nucleobase within a nucleotide in the AStD. In some embodiments, the ASGPR-binding moiety is present within an internucleotide bond in the AStD. In some embodiments, the ASGPR-binding moiety is present on a terminus (e.g., the 5' or 3' terminus) within the AStD.

[0017] In some embodiments, ASt domain 1 comprises positions 1-9 within the TREM sequence. In some embodiments, the ASGPR-binding moiety is present within ASt domain 1 (e.g., positions 1-9) within the TREM sequence. In some embodiments, ASt domain 2 comprises positions 65-76 within the TREM sequence. In some embodiments, the ASGPR-binding moiety is present within ASt domain 1 (e.g., positions 65-76) within the TREM sequence.

[0018] In some embodiments, the AStD is located below the corresponding sequence of the consensus sequence provided in the "consensus sequence" section, or differs from the consensus sequence at no more than 1, 2, 5, or 10 positions. In some embodiments, the ASPGR binding moiety is located below the corresponding sequence of the consensus sequence provided in the "consensus sequence" section, or with an AStD that differs from the consensus sequence at no more than 1, 2, 5, or 10 positions.

[0019] In one embodiment, AStD has formula I ZZZ Residues R1-R2-R3-R4-R5-R6-R7 (exemplary ASt domain 2) and residues R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 (exemplary ASt domain 2), where ZZZ represents any of the 20 amino acids. In some embodiments, ZZZ refers to all species.

[0020] In one embodiment, AStD has formula II ZZZ Residues R1-R2-R3-R4-R5-R6-R7 and residues R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 where ZZZ represents any of 20 amino acids.

[0021] In some embodiments, Formula II ZZZrefers to a mammal.

[0022] In certain embodiments, AStD has formula III ZZZ Residues R1-R2-R3-R4-R5-R6-R7 and residues R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 In some embodiments, the amino acid sequence of Formula III is ZZZ refers to humans.

[0023] In one embodiment, ZZZ represents any of the amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.

[0024] "Anticodon hairpin domain (ACHD)," as that term is used herein, refers to a domain containing an anticodon that binds to a respective codon in an mRNA, e.g., containing a sufficient sequence, e.g., an anticodon triplet, to mediate pairing with the codon (with or without wobble) when present in an otherwise wild-type tRNA. In some embodiments, the ACHD has at least 75, 80, 85, 85, 90, 95, or 100% identity to a naturally occurring ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1. In some embodiments, a TREM may comprise a fragment or analog of an ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1, wherein the fragment in some embodiments has ACHD activity and in other embodiments does not have ACHD activity. In some embodiments, an ASGPR-binding moiety is present within an ACHD. In some embodiments, the ASGPR-binding moiety is bound to a nucleobase within a nucleotide in the ACHD.

[0025] In some embodiments, the ACHD comprises positions 27 to 43 within the TREM sequence. In some embodiments, the ASGPR binding moiety is present within the ACHD (eg, positions 27 to 43) within the TREM sequence.

[0026] In some embodiments, the ACHD is located below the sequence corresponding to the consensus sequence provided in the "consensus sequence" section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions. In some embodiments, the ASGPR-binding moiety is located below the sequence corresponding to the consensus sequence provided in the "consensus sequence" section, or within a sequence that differs from the consensus sequence by no more than 1, 2, 5, or 10 positions.

[0027] In one embodiment, ACHD has formula I ZZZ Residue -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 In some embodiments, the amino acid sequence of Formula I is ZZZ refers to all species.

[0028] In one embodiment, ACHD has formula II ZZZ Residue -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R45 -R 46 In some embodiments, the amino acid sequence of Formula II is ZZZ refers to a mammal.

[0029] In certain embodiments, ACHD has formula III ZZZ Residue -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 In some embodiments, the amino acid sequence of Formula III is ZZZ refers to humans.

[0030] In one embodiment, ZZZ represents any of the amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.

[0031] In some embodiments, the anticodon of the TREM entity comprises three nucleotide residues and pairs with a three nucleotide codon. In some embodiments, the anticodon of the TREM entity consists of three nucleotide residues and pairs with an anticodon consisting of three nucleotide residues. In some embodiments, the anticodon of the TREM entity does not pair with codons having four, five or more nucleotide residues, but only pairs with three codon nucleotide residues.

[0032] In certain embodiments, the TREM entity does not alter the reading frame of the mRNA, hi certain embodiments, the anticodon of the TREM entity pairs with the triplet codon of the mRNA and does not pair with the adjacent nucleotide.

[0033] In some embodiments, the use of a TREM entity does not alter the length of the polypeptide transcribed from the mRNA, e.g., it does not suppress a stop codon, e.g., a premature stop codon. In some embodiments, the TREM does not alter the length of the ORF of the mRNA.

[0034] "Asialoglycoprotein receptor (ASGPR) binding moiety," as that term is used herein, refers to a moiety that binds to an asialoglycoprotein receptor. In certain embodiments, the ASGPR binding moieties described herein refer to a structure comprising (i) an ASGPR carbohydrate and (ii) an ASGPR linker (e.g., a linker connecting the carbohydrate to a TREM). Exemplary ASGPR moieties include a galactose (Gal), galactosamine (GalNH), or N-acetylgalactosamine (GalNAc) moiety, such as Gal, GalNH, or GalNAc, or analogs thereof. The ASGPR binding moiety may include a functional group (e.g., a hydroxyl group, a carboxylic acid group, an amine) that can be protected by a chemical protecting group, e.g., an acetyl group or a methyl group. In certain embodiments, the ASGPR binding moiety includes a tri-antennary GalNAc moiety. In certain embodiments, the ASGPR binding moiety may be an ASGPR binding moiety described in further detail herein.

[0035] A "cognate adaptor function TREM," as that term is used herein, refers to a TREM that mediates initiation or elongation by the AA that is naturally associated with the anticodon of the TREM (the cognate AA).

[0036] "Decreased expression," as that term is used herein, refers to a decrease compared to a reference, for example, if modification of a control region or addition of an agent results in decreased expression of the product of interest, which is decreased compared to other similar cells without the modification or addition.

[0037] Dihydrouridine hairpin domain (DHD), as that term is used herein, refers to a domain that, when present in, for example, an otherwise wild-type tRNA, mediates recognition by an aminoacyl-tRNA synthetase, e.g., comprises a sufficient RNA sequence to act as a recognition site for an aminoacyl-tRNA synthetase for amino acid loading of a TREM. In embodiments, the DHD mediates stabilization of the tertiary structure of a TREM. In some embodiments, the DHD has at least 75, 80, 85, 85, 90, 95, or 100% identity to a naturally occurring DHD, e.g., a DHD encoded by a nucleic acid in Table 1. In some embodiments, a TREM may comprise a fragment or analog of a DHD, e.g., a DHD encoded by a nucleic acid in Table 1, wherein the fragment in some embodiments has DHD activity and in other embodiments does not have DHD activity. In some embodiments, an ASGPR-binding moiety is present within the DHD. In some embodiments, the ASGPR-binding moiety is attached to a nucleobase within a nucleotide in the DHD.

[0038] In some embodiments, the DHD comprises positions 10 to 26 within the TREM sequence. In some embodiments, the ASGPR binding moiety is present within the DHD (eg, positions 10 to 26) within the TREM sequence.

[0039] In some embodiments, the DHD is located below the sequence corresponding to the consensus sequence provided in the "consensus sequence" section, or differs from the consensus sequence at no more than 1, 2, 5, or 10 positions. In some embodiments, the ASGPR-binding moiety is located below the sequence corresponding to the consensus sequence provided in the "consensus sequence" section, or within a sequence that differs from the consensus sequence at no more than 1, 2, 5, or 10 positions.

[0040] In certain embodiments, the DHD has formula I ZZZ Residue R of 10 -R 11 -R 12 -R 13 -R 14 R 15 -R16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 In some embodiments, the amino acid sequence of Formula I is ZZZ refers to all species.

[0041] In certain embodiments, the DHD has Formula II ZZZ Residue R of 10 -R 11 -R 12 -R 13 -R 14 R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 In some embodiments, the amino acid sequence of Formula II is ZZZ refers to a mammal.

[0042] In certain embodiments, the DHD has formula III ZZZ Residue R of 10 -R 11 -R 12 -R 13 -R 14 R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R27 -R 28 In some embodiments, the amino acid sequence of Formula III is ZZZ refers to humans.

[0043] In one embodiment, ZZZ represents any of the amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.

[0044] "Foreign nucleic acid," as that term is used herein, refers to a nucleic acid sequence that is not present in, or differs by at least one nucleotide from, the closest sequence in a reference cell, e.g., the cell into which the foreign nucleic acid is introduced. In one embodiment, the foreign nucleic acid comprises a nucleic acid encoding a TREM.

[0045] "Extraneous TREM," as that term is used herein, means: (a) differs in at least one nucleotide or one post-transcriptional modification from the closest sequence tRNA in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced; (b) whether it has been introduced into cells other than those in which it was transcribed; (c) it is present in cells other than those in which it naturally occurs; or (d) A TREM that has an expression profile, e.g., a level or distribution, that is not wild-type (e.g., it is expressed at a level higher than wild-type). In some embodiments, the expression profile can be mediated by changes introduced into a nucleic acid that regulates expression or by the addition of an agent that regulates expression of an RNA molecule. In some embodiments, an exogenous TREM comprises one, two, three, or four of characteristics (a)-(d).

[0046] "GMP-grade composition," as that term is used herein, refers to a composition that complies with current Good Manufacturing Practice (cGMP) guidelines or other similar requirements. In certain embodiments, a GMP-grade composition may be used as a pharmaceutical product.

[0047] As used herein, the terms "increase" and "decrease" refer to modulation that results in an increase or decrease, respectively, in the amount of a particular indicator of function, expression, or activity relative to a reference. For example, after administration of a TREM as described herein to a cell, tissue, or subject, the amount of a marker of the indicator as described herein (e.g., protein translation, mRNA stability, protein folding) can be increased or decreased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%, 2-fold, 3-fold, 5-fold, 10-fold, or more relative to the amount of the marker before administration or relative to the effect of a negative control agent. The indicator can be measured after administration at a time when administration has had the recited effect, for example, at least 12 hours, 24 hours, 1 week, 1 month, 3 months, or 6 months after treatment begins.

[0048] "Increased expression," as that term is used herein, refers to an increase compared to a reference, for example, if modification of a control region or addition of an agent results in increased expression of the product of interest, which is increased compared to an otherwise similar cell without the modification or addition.

[0049] Linker 2 region (L2), as that term is used herein, refers to the linker that includes residues R8-R9 of the consensus sequence provided in the "consensus sequence" section.

[0050] The linker 3 region (L3), as that term is used herein, is a region consisting of residues R of the consensus sequence provided in the "consensus sequence" section. 29 refers to a linker comprising:

[0051] The "linker 4 region (L4), as that term is used herein, is defined as the residues R of the consensus sequence provided in the "consensus sequence" section. 72 Refers to a domain that includes

[0052] "Modification," as the term is used herein with respect to a nucleotide, refers to a modification of the chemical structure, e.g., a covalent modification of the subject nucleotide. In some embodiments, the modification is present in the nucleobase, nucleotide sugar, or internucleotide linkage of the nucleotide of the TREM. The modification can be natural or non-natural. In some embodiments, the modification is non-natural. In some embodiments, the modification is natural. In some embodiments, the modification is a synthetic modification. In some embodiments, the modification is a modification shown in Table 5, 6, 7, 8, or 9.

[0053] A "naturally occurring nucleotide," as that term is used herein, refers to a nucleotide that does not contain a non-naturally occurring modification. In some embodiments, it contains a naturally occurring modification.

[0054] "Nucleotide," as that term is used herein, refers to an entity that includes a sugar, typically a pentameric sugar; a nucleobase; and a phosphate linkage group (e.g., an internucleotide linkage). In some embodiments, a nucleotide includes a naturally occurring nucleotide, e.g., a naturally occurring nucleotide in a human cell, such as an adenine, thymine, guanine, cytosine, or uracil nucleotide.

[0055] "Thymine hairpin domain (THD), as that term is used herein, refers to a domain that comprises sufficient RNA sequence to mediate ribosome recognition, e.g., when present in an otherwise wild-type tRNA, to act as a recognition site for the ribosome to form a TREM-ribosome complex during translation. In some embodiments, the THD is at least 75, 80, 85, 85, 90, 95, or 100% identical to a naturally occurring THD, e.g., a THD encoded by a nucleic acid in Table 1. In some embodiments, a TREM may comprise a fragment or analog of a THD, e.g., a THD encoded by a nucleic acid in Table 1, wherein the fragment in embodiments has THD activity and in other embodiments does not have THD activity. In some embodiments, an ASGPR-binding moiety is present in a THD. In some embodiments, an ASGPR-binding moiety is attached to a nucleobase within a nucleotide in a THD.

[0056] In some embodiments, the THD comprises positions 50 to 64 within the TREM sequence. In some embodiments, the ASPGR binding moiety is present within the THD (eg, positions 50 to 64) within the TREM sequence.

[0057] In one embodiment, the THD is below the corresponding sequence of the consensus sequence provided in the "consensus sequence" section or differs from the consensus sequence at no more than 1, 2, 5, or 10 positions.

[0058] In certain embodiments, THD has formula I ZZZ Residue -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64In some embodiments, the amino acid sequence of Formula I is ZZZ refers to all species.

[0059] In certain embodiments, THD has the formula II ZZZ Residue -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 In some embodiments, the amino acid sequence of Formula II is ZZZ refers to a mammal.

[0060] In certain embodiments, THD has the formula II ZZZ Residue -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 In some embodiments, the amino acid sequence of Formula III is ZZZ refers to humans.

[0061] In one embodiment, ZZZ represents any of the amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.

[0062] "tRNA-based effector molecule" or "TREM," as that term is used herein, refers to an RNA molecule comprising the structure or characteristics of (a)-(v) below, which may be a recombinant TREM, a synthetic TREM, or a TREM expressed from a heterologous cell. The TREMs described in this invention are synthetic molecules, e.g., made in a cell-free reaction, e.g., a solid-phase or solution-phase synthesis reaction. TREMs are chemically distinct, e.g., in terms of primary sequence, type, or location of modification, from endogenous tRNA molecules made in a cell, e.g., a mammalian cell, e.g., a human cell. TREMs may have the structure and function of more than one (e.g., 2, 3, 4, 5, 6, 7, 8, 9) of (a)-(v).

[0063] In some embodiments, a TREM is chemical, as assessed by its structure or the way in which it is made.

[0064] In certain embodiments, the TREM comprises one or more of the following structures or characteristics: (a') an optional linker region of the consensus sequence provided in the "consensus sequence" section, e.g., the linker 1 region; (a) acceptor stem domain (AStD), which typically contains ASt domain 1 and ASt domain 2; (a'-1) linker 2 region (L2) (a linker comprising residues R8 to R9 of the consensus sequence provided in the "consensus sequence" section), e.g., linker 2 region; (b) DHD or dihydrouridine hairpin domain (DHD); (b'-1) linker 3 region, or L3; (c) ACHD or anticodon hairpin domain; (d) VLD, or variable loop domain (VLD); (e) THD or thymine hairpin domain (THD); (e'1) Residue R of the consensus sequence provided in the "consensus sequence" section 72 an L4 linker comprising: (f) under physiological conditions, it comprises a stem structure and one or more loop structures, e.g., one, two, or three loops. The loop may comprise a domain described herein, e.g., a domain selected from (a)-(e). The loop may comprise one or more domains. In some embodiments, the stem or loop structure has at least 75, 80, 85, 85, 90, 95, or 100% identity to a naturally occurring stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1. In some embodiments, the TREM may comprise a fragment or analog of a stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1, where the fragment in embodiments has the activity of the stem or loop structure and in other embodiments does not have the activity of the stem or loop structure; (g) tertiary structure, e.g., L-type tertiary structure; (h) Adaptor function, i.e., TREM, mediates the acceptance of an amino acid, e.g., its cognate amino acid, and mediates the transfer of AA in the initiation or elongation of a polypeptide chain; (i) cognate adaptor function (TREMs mediate acceptance and incorporation of amino acids (e.g., cognate amino acids) that are naturally linked to the anticodon of the TREM initiating or elongating a polypeptide chain); (j) noncognate adaptor function (TREM mediates the acceptance and incorporation of amino acids other than those naturally linked to the anticodon of the TREM (e.g., noncognate amino acids) in the initiation or elongation of a polypeptide chain); (k) a regulatory function, such as an epigenetic function (e.g., a gene silencing function or a signal pathway regulation function), a cell fate regulation function, an mRNA stability regulation function, a protein stability regulation function, a protein transduction regulation function, or a protein compartmentalization function; (l) structures that allow ribosome binding; (m) post-transcriptional modifications, e.g., naturally occurring post-transcriptional modifications; (n) a functional property of tRNA, e.g., the ability to inhibit any of the properties (h)-(k) possessed by tRNA; (o) the ability to regulate cell fate; (p) the ability to regulate ribosome occupancy; (q) the ability to regulate protein translation; (r) the ability to regulate mRNA stability; (s) the ability to regulate protein folding and structure; (t) the ability to regulate protein transduction or compartmentalization; (u) the ability to modulate protein stability; or (v) The ability to modulate a signaling pathway, for example, a cellular signaling pathway.

[0065] In certain embodiments, the TREM comprises a full-length tRNA molecule or a fragment thereof.

[0066] In one embodiment, the TREM comprises the following properties: (a) through (e).

[0067] In one embodiment, the TREM comprises the following properties: (a) and (c).

[0068] In one embodiment, the TREM comprises the following characteristics: (a), (c), and (h).

[0069] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), and (b).

[0070] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), and (e).

[0071] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (b), and (e).

[0072] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (b), (e), and (g).

[0073] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), and (m).

[0074] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), and (g).

[0075] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), and (b).

[0076] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), and (e).

[0077] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), (g), (b), and (e).

[0078] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), (g), (b), (e), and (q).

[0079] In one embodiment, the TREM is (i) an amino acid binding domain that binds to an amino acid (e.g., an AStD as described in (a) herein); and (ii) It contains an anticodon (e.g., ACHD as described in (c) herein) that binds to each codon in the mRNA.

[0080] In certain embodiments, the TREM comprises a flexible RNA linker that provides the covalent bond between (i) and (ii).

[0081] In certain embodiments, TREM mediates protein translation.

[0082] In some embodiments, the TREM comprises a linker, e.g., an RNA linker, e.g., a flexible RNA linker, that provides a covalent bond between the first and second structures or domains. In some embodiments, the RNA linker comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ribonucleotides. The TREM may comprise one or more linkers; for example, in embodiments, a TREM comprising (a), (b), (c), (d), and (e) may have a first linker between the first and second domains and a second linker between the third domain and another domain.

[0083] In one embodiment, the TREM comprises the sequence of Formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2].

[0084] In some embodiments, a TREM comprises an RNA sequence that is at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical to, or differs by no more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 ribonucleotides from, an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In some embodiments, a TREM comprises an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In some embodiments, a TREM comprises an RNA sequence encoded by a DNA sequence that is at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical to, a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In some embodiments, the TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof, that is at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical to, or differs by no more than 1, 2, 3, 4, 5, 10, or 15 ribonucleotides therefrom. In some embodiments, the TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In some embodiments, the TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by a DNA sequence that is at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical to, a DNA sequence listed in Table 1, or a fragment or functional fragment thereof.

[0085] In certain embodiments, the TREM is 76 to 90 nucleotides in length. In certain embodiments, the TREM, or a fragment or functional fragment thereof, is 10 to 90 nucleotides, 10 to 80 nucleotides, 10 to 70 nucleotides, 10 to 60 nucleotides, 10 to 50 nucleotides, 10 to 40 nucleotides, 10 to 30 nucleotides, 10 to 20 nucleotides, 20 to 90 nucleotides, 20 to 80 nucleotides, 20 to 70 nucleotides, 20 to 60 nucleotides, 20 to 50 nucleotides, 20 to 40 nucleotides, 30 to 90 nucleotides, 30 to 80 nucleotides, 30 to 70 nucleotides, 30 to 60 nucleotides, or 30 to 50 nucleotides.

[0086] In certain embodiments, the TREM is aminoacylated, eg, charged with an amino acid by an aminoacyl-tRNA synthetase.

[0087] In certain embodiments, the TREM is not loaded with amino acids, eg, unloaded TREM (uTREM).

[0088] In some embodiments, a TREM comprises a less than full-length tRNA. In embodiments, a TREM can correspond to a naturally occurring or non-naturally occurring fragment of a tRNA. Exemplary fragments include a TREM half (e.g., derived from an ACHD, e.g., a cleavage in the anticodon sequence, e.g., the 5' half or the 3' half); a 5' fragment (e.g., derived from a cleavage in the DHD or ACHD, e.g., a fragment including the 5' end); a 3' fragment (e.g., derived from a cleavage in the THD, e.g., a fragment including the 3' end); or an internal fragment (e.g., derived from a cleavage in one or more of the ACHD, DHD, or THD).

[0089] A "TREM core fragment," as that term is used herein, is a fragment having the sequence of Formula B: [L1] y -[ASt domain 1] x -[L2] y -[DH domain] y -[L3] y -[ACH domain] x-[VL domain] y -[TH domain] y -[L4] y -[ASt domain 2] x where x=1 and y=0 or 1.

[0090] "TREM fragment," as used herein, refers to a portion of TREM, wherein the TREM comprises the sequence of Formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2].

[0091] A "non-cognate adapter function TREM," as that term is used herein, refers to a TREM that mediates initiation or extension by an AA other than the AA naturally associated with the anticodon of the TREM (non-cognate AA). In one embodiment, a non-cognate adapter function TREM is also referred to as a mischarged TREM (mTREM).

[0092] A "non-naturally occurring sequence," as that term is used herein, refers to a sequence in which adenine is replaced by a residue other than an analog of adenine, cytosine is replaced by a residue other than an analog of cytosine, guanine is replaced by a residue other than an analog of guanine, and uracil is replaced by a residue other than an analog of uracil. An analog refers to any possible derivative of ribonucleotides A, G, C, or U. In one embodiment, a sequence having a derivative of any one of ribonucleotides A, G, C, or U is a non-naturally occurring sequence.

[0093] "Pharmaceutical TREM composition," as that term is used herein, refers to a TREM composition suitable for pharmaceutical use. Typically, a pharmaceutical TREM composition includes a pharmaceutical excipient. In some embodiments, the TREM will be the only active ingredient in a pharmaceutical TREM composition. In embodiments, a pharmaceutical TREM composition is free, substantially free, or has less than a pharmaceutically acceptable amount of host cell proteins, DNA, e.g., host cell DNA, endotoxins, and bacteria.

[0094] "Post-transcriptional processing," as the term is used herein with respect to a molecule of interest, e.g., a TREM, RNA, or tRNA, refers to the covalent modification of the molecule of interest. In some embodiments, the covalent modification occurs post-transcriptionally. In some embodiments, the covalent modification occurs co-transcriptionally. In some embodiments, the modification is made in vivo, e.g., in the cells used to generate the TREM. In some embodiments, the modification is made ex vivo, e.g., on a TREM isolated or obtained from the cells that generated the TREM. In some embodiments, the post-transcriptional modification is selected from the post-transcriptional modifications listed in Table 2.

[0095] "Subject," as the term is used herein, includes any living organism, such as a human or other animal. In embodiments, the subject is a vertebrate (e.g., a mammal, a bird, a fish, a reptile, or an amphibian). In embodiments, the subject is a mammal, e.g., a human. In embodiments, the subject of the present methods is a non-human mammal. In embodiments, the subject is a non-human mammal, such as a non-human primate (e.g., a monkey, an ape), an ungulate (e.g., a cow, a buffalo, a sheep, a goat, a pig, a camel, a llama, an alpaca, a deer, a horse, a donkey), a carnivore (e.g., a dog, a cat), a rodent (e.g., a rat, a mouse), or a lagomorph (e.g., a rabbit). In embodiments, the subject is a bird, such as a member of the avian taxon Galliformes (e.g., chicken, turkey, pheasant, quail), Anseriformes (e.g., duck, goose), Paleognatha (e.g., ostrich, emu), Columbiformes (e.g., pigeon, dove), or Psittaciformes (e.g., parrot). The subject can be male or female of any age, e.g., a pediatric subject (e.g., infant, toddler, adolescent) or an adult subject (e.g., young adult, middle-aged adult, or elderly). The non-human subject can be a transgenic animal.

[0096] "Synthetic TREM," as that term is used herein, refers to a TREM synthesized outside of or by a cell that has endogenous nucleic acid encoding the TREM; for example, a synthetic TREM is synthesized by cell-free solid-phase synthesis. A synthetic TREM may have the same or a different sequence or tertiary structure as a naturally occurring tRNA.

[0097] "Recombinant TREM," as that term is used herein, refers to a TREM expressed in a cell that has been modified by human intervention, with modifications that mediate the production of the TREM (e.g., the cell includes an exogenous sequence encoding the TREM), or modifications that mediate expression, e.g., transcriptional expression or post-transcriptional modification, of the TREM. A recombinant TREM may have the same or a different sequence, set of post-transcriptional modifications, or tertiary structure as a reference tRNA, e.g., a naturally occurring tRNA.

[0098] "tRNA," as the term is used herein, refers to a naturally occurring transfer ribonucleic acid in its natural state.

[0099]

[0023] A "TREM composition," as that term is used herein, refers to a composition comprising multiple TREMs, multiple TREM core fragments, and / or multiple TREM fragments. A TREM composition may comprise one or more species of TREM, TREM core fragment, or TREM fragment. In some embodiments, a composition comprises only a single species of TREM, TREM core fragment, or TREM fragment. In some embodiments, a TREM composition comprises a first TREM, TREM core fragment, or TREM fragment species; and a second TREM, TREM core fragment, or TREM fragment species. In some embodiments, a TREM composition comprises X TREM, TREM core fragment, or TREM fragment species (X=2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, a TREM, TREM core fragment, or TREM fragment has at least 70, 75, 80, 85, 90, or 95, or 100% identity to a sequence encoded by a nucleic acid in Table 1. A TREM composition may comprise one or more species of TREM, TREM core fragment, or TREM fragment. In some embodiments, the TREM composition is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or 99% dry weight TREM (for liquid compositions, dry weight refers to the weight after removal of substantially all liquid, e.g., after lyophilization). In some embodiments, the composition is a liquid. In some embodiments, the composition is a dry, e.g., lyophilized, material. In some embodiments, the composition is a frozen composition. In some embodiments, the composition is sterile. In some embodiments, the composition comprises at least 0.5 g, 1.0 g, 5.0 g, 10 g, 15 g, 25 g, 50 g, 100 g, 200 g, 400 g, or 500 g (e.g., as measured by dry weight) TREM.

[0100] In some embodiments, at least X% of the TREMs in the TREM composition comprise a chemical modification at a selected position, where X is 80, 90, 95, 96, 97, 98, 99, or 99.5.

[0101] In some embodiments, at least X% of the TREMs in the TREM composition comprise a chemical modification at a first position and a chemical modification at a second position, where X is independently 80, 90, 95, 96, 97, 98, 99, or 99.5. In some embodiments, the modifications at the first and second positions are the same. In some embodiments, the modifications at the first and second positions are different. In some embodiments, the nucleotides at the first and second positions are the same, e.g., both adenine. In some embodiments, the nucleotides at the first and second positions are different, e.g., one adenine and one thymine.

[0102] In some embodiments, at least X% of the TREMs in a TREM composition comprise a chemical modification at a first position and less than Y% have a chemical modification at a second position, where X is 80, 90, 95, 96, 97, 98, 99, or 99.5 and Y is 20, 20, 5, 2, 1, 0.1, or 0.01. In some embodiments, the nucleotides at the first and second positions are the same, e.g., both adenine. In some embodiments, the nucleotides at the first and second positions are different, e.g., one adenine and one thymine.

[0103] "Variable loop domain (VLD)," as that term is used herein, refers to a domain that comprises sufficient RNA sequence to mediate aminoacyl-tRNA synthetase recognition, e.g., to act as a recognition site for an aminoacyl-tRNA synthetase for amino acid charging of a TREM, e.g., when present in an otherwise wild-type tRNA. In embodiments, the VLD mediates stabilization of the tertiary structure of a TREM. In some embodiments, the VLD modulates, e.g., increases, the specificity of a TREM for its cognate amino acid, e.g., the VLD modulates the cognate adaptor function of a TREM. In some embodiments, the VLD has at least 75, 80, 85, 85, 90, 95, or 100% identity to a naturally occurring VLD, e.g., a VLD encoded by a nucleic acid in Table 1. In some embodiments, a TREM may comprise a fragment or analog of a VLD, e.g., a VLD encoded by a nucleic acid in Table 1, wherein the fragment in embodiments has VLD activity and in other embodiments does not have VLD activity. In certain embodiments, the ASGPR binding moiety is present within a VLD. In certain embodiments, the ASGPR binding moiety is attached to a nucleobase within a nucleotide in the VLD.

[0104] In some embodiments, the VLD comprises positions 44 to 49 within the TREM sequence. In some embodiments, the ASGPR binding moiety is present within the VLD (eg, positions 44 to 49) within the TREM sequence.

[0105] In certain embodiments, the VLD is below the corresponding sequence of the consensus sequence provided in the "consensus sequence" section.

[0106] In one embodiment, the VLD is a residue of the consensus sequence provided in the "consensus sequence" section -[R 47 ] xx=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18 , x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271).

[0107] TREM Entity Described herein are TREM entities, such as TREMs, TREM core fragments, or TREM fragments, modified with asialoglycoprotein receptor (ASGPR) binding moieties, as well as compositions and methods of use thereof. TREM entities (e.g., TREMs) refer to RNA molecules that contain one or more of the properties described herein. ASGPR binding moieties can be conjugated to nucleobases within the TREM entity, within the internucleotide bond of the TREM entity, or at the end (e.g., 5' or 3' end) of the TREM entity. TREM entities (e.g., TREMs) can include, for example, chemical modifications such as those provided in Tables 4, 5, 6, or 7.

[0108] In certain embodiments, the TREM entity comprises a TREM comprising the sequence of formula A; a TREM core fragment comprising the sequence of formula B; or a TREM fragment comprising a portion of the TREM comprising the sequence of formula A.

[0109] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR-binding moiety is present within ASt domain 1 (e.g., on a nucleobase, at the end (e.g., the 5' end), or within an internucleotide linkage of ASt domain 1). In some embodiments, the ASGPR-binding moiety is present on a nucleobase of a nucleotide within ASt domain 1. In some embodiments, the ASGPR-binding moiety is present at the 5' end or at [L1] within ASt domain 1. In some embodiments, the ASGPR-binding moiety is present within an internucleotide linkage of ASt domain 1. In some embodiments, [VL domain] is optional. In some embodiments, [L1] is optional.

[0110] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR-binding moiety is present within ASt domain 2 (e.g., on a nucleobase, at the terminus (e.g., the 3' terminus), or within an internucleotide linkage of ASt domain 2). In some embodiments, the ASGPR-binding moiety is present on a nucleobase of a nucleotide within ASt domain 2. In some embodiments, the ASGPR-binding moiety is present at the 3' terminus within ASt domain 2. In some embodiments, the ASGPR-binding moiety is present within an internucleotide linkage of ASt domain 2. In some embodiments, [VL domain] is optional. In some embodiments, [L1] is optional.

[0111] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR-binding moiety is present within one or both of ASt domain 1 and ASt domain 2 (e.g., on a nucleobase, at the terminus (e.g., the 5' or 3' terminus), or within an internucleotide linkage of ASt domain 1 or ASt domain 2). In some embodiments, the ASGPR-binding moiety is present on a nucleobase of a nucleotide within ASt domain 1 or ASt domain 2. In some embodiments, the ASGPR-binding moiety is present at the 5' terminus or [L1] within ASt domain 1 or the 3' terminus within ASt domain 2. In some embodiments, the ASGPR-binding moiety is present within an internucleotide linkage of ASt domain 1 or ASt domain 2. In some embodiments, [VL domain] is optional. In some embodiments, [L1] is optional.

[0112] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR binding moiety is present within the DH domain (e.g., on a nucleobase or within an internucleotide linkage of the DH domain). In some embodiments, the ASGPR binding moiety is present on a nucleobase of a nucleotide within the DH domain. In some embodiments, the ASGPR binding moiety is present within an internucleotide linkage of the DH domain. In some embodiments, [L1] is optional.

[0113] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR binding moiety is within the ACH domain (e.g., on a nucleobase or within an internucleotide linkage of the ACH domain). In some embodiments, the ASGPR binding moiety is present on a nucleobase of a nucleotide within the ACH domain. In some embodiments, the ASGPR binding moiety is present within an internucleotide linkage of the ACH domain. In some embodiments, [VL domain] is optional. In some embodiments, [L1] is optional.

[0114] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR binding moiety is present within the VL domain (e.g., on a nucleobase or within an internucleotide linkage of the VL domain). In some embodiments, the ASGPR binding moiety is present on a nucleobase of a nucleotide within the VL domain. In some embodiments, the ASGPR binding moiety is present within an internucleotide linkage of the VL domain. In some embodiments, [L1] is optional.

[0115] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR binding moiety is present within the TH domain (e.g., on a nucleobase or within an internucleotide linkage of the TH domain). In some embodiments, the ASGPR binding moiety is present on a nucleobase of a nucleotide within the TH domain. In some embodiments, the ASGPR binding moiety is present within an internucleotide linkage of the TH domain. In some embodiments, [VL domain] is optional. In some embodiments, [L1] is optional.

[0116] In some embodiments, the TREM comprises a sequence of Formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR binding moiety is bound to a nucleobase in one or more domains selected from [ASt domain 1], [DH domain], [ACH domain], [TH domain], and / or [ASt domain 2]. In some embodiments, the [VL domain] is optional. In some embodiments, [L1] is optional.

[0117] In some embodiments, the TREM comprises a sequence of formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the ASGPR-binding moiety is attached to an internucleotide linkage within one or more domains selected from [ASt domain 1], [DH domain], [ACH domain], [TH domain], and / or [ASt domain 2]. In some embodiments, [VL domain] is optional. In some embodiments, [L1] is optional.

[0118] In one embodiment, the TREM core fragment has the formula B:[L1] y -[ASt domain 1] x -[L2] y -[DH domain] y -[L3] y -[ACH domain] x -[VL domain] y -[TH domain] y -[L4] y -[ASt domain 2] x wherein x=1 and y=0 or 1, and the ASGPR-binding moiety is attached to a nucleobase within a nucleotide within one or both of ASt domain 1 and ASt domain 2. In some embodiments, y=0. In some embodiments, y=1.

[0119] In one embodiment, the TREM core fragment has the formula B:[L1] y-[ASt domain 1] x -[L2] y -[DH domain] y -[L3] y -[ACH domain] x -[VL domain] y -[TH domain] y -[L4] y -[ASt domain 2] x wherein x=1 and y=0 or 1, and the ASGPR-binding moiety is attached to a nucleobase within a nucleotide within the DH domain. In some embodiments, y=0. In some embodiments, y=1.

[0120] In one embodiment, the TREM core fragment has the formula B:[L1] y -[ASt domain 1] x -[L2] y -[DH domain] y -[L3] y -[ACH domain] x -[VL domain] y -[TH domain] y -[L4] y -[ASt domain 2] x wherein x=1 and y=0 or 1, and the ASGPR-binding moiety is attached to a nucleobase within a nucleotide within the ACH domain. In some embodiments, y=0. In some embodiments, y=1.

[0121] In one embodiment, the TREM core fragment has the formula B:[L1] y -[ASt domain 1] x -[L2] y -[DH domain] y -[L3] y -[ACH domain] x -[VL domain] y -[TH domain] y -[L4] y -[ASt domain 2] xwherein x=1 and y=0 or 1, and the ASGPR-binding moiety is attached to a nucleobase within a nucleotide in the TH domain. In some embodiments, y=0. In some embodiments, y=1.

[0122] In one embodiment, the TREM core fragment has the formula B:[L1] y -[ASt domain 1] x -[L2] y -[DH domain] y -[L3] y -[ACH domain] x -[VL domain] y -[TH domain] y -[L4] y -[ASt domain 2] x wherein x=1 and y=0 or 1, and the ASGPR-binding moiety is bound to a nucleobase within one or more domains selected from [ASt domain 1], [DH domain], [ACH domain], [TH domain], and / or [ASt domain 2]. In some embodiments, y=0. In some embodiments, y=1.

[0123] In one embodiment, a TREM fragment comprises a portion of TREM, wherein the TREM comprises the sequence of Formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], and the TREM fragment comprises one, two, three, or all, or any combination, of the following: a TREM half (e.g., derived from a cleavage in the ACH domain, e.g., at the anticodon sequence, e.g., the 5' half or the 3' half); a 5' fragment (e.g., derived from a cleavage in the DH domain or ACH domain, e.g., a fragment including the 5' end); a 3' fragment (e.g., derived from a cleavage in the TH domain, e.g., a fragment including the 3' end); or an internal fragment (e.g., derived from a cleavage in any one of the ACH domain, DH domain, or TH domain). Exemplary TREM fragments include a TREM half (e.g., derived from a cleavage in the ACHD, e.g., a 5' TREM half or a 3' TREM half), a 5' fragment (e.g., derived from a cleavage in the DHD or ACHD, e.g., a fragment including the 5' end), a 3' fragment (e.g., derived from a cleavage in the THD, e.g., a fragment including the 3' end of the TREM), or an internal fragment (e.g., derived from a cleavage in one or more of the ACHD, DHD, or THD).

[0124] In some embodiments, the TREM, TREM core fragment, or TREM fragment may be charged with an amino acid (e.g., a cognate amino acid); may be charged with a non-cognate amino acid (e.g., a misloaded TREM (mTREM)); or may not be charged with an amino acid (e.g., an unloaded TREM (uTREM)). In some embodiments, the TREM, TREM core fragment, or TREM fragment may be charged with an amino acid selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.

[0125] In some embodiments, the TREM, TREM core fragment or TREM fragment is a cognate TREM. In some embodiments, the TREM, TREM core fragment or TREM fragment is a non-cognate TREM. In some embodiments, the TREM, TREM core fragment or TREM fragment recognizes a codon provided in Table 2 or Table 3.

[0126] [Table 1]

[0127] [Table 2]

[0128] In some embodiments, the TREM comprises a deoxyribonucleic acid (DNA) sequence disclosed in Table 1, e.g., a ribonucleic acid (RNA) sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In some embodiments, the TREM comprises an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM comprises an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0129] In some embodiments, the TREM, TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 contiguous nucleotides of an RNA sequence encoded by a DNA sequence disclosed in Table 1, for example, at least 5, 10, 15, 20, 25, or 30 contiguous nucleotides of an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In some embodiments, the TREM, TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 contiguous nucleotides of an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, for example, an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM, TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, for example, any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0130] In one embodiment, the TREM core fragment or TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the RNA sequence encoded by the DNA sequence provided in Table 1, for example, any one of SEQ ID NOs: 1 to 451 disclosed in Table 1. In one embodiment, the TREM core fragment or TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a DNA sequence provided in Table 1, for example, an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM core fragment or TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a DNA sequence provided in Table 1, for example, any one of SEQ ID NOs: 1 to 451 disclosed in Table 1.

[0131] In one embodiment, the TREM core fragment or TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt, or 60 nt (but less than the full length) of a DNA sequence disclosed in Table 1, for example, an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM core fragment or TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt, or 60 nt (but less than the full length) of an RNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a DNA sequence provided in Table 1, for example, an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM core fragment or TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence having at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identity to a DNA sequence provided in Table 1, for example, any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0132] In one embodiment, the TREM core fragment or TREM fragment comprises a sequence of 10 to 90 ribonucleotides (rnt), 10 to 80rnt, 10 to 70rnt, 10 to 60rnt, 10 to 50rnt, 10 to 40rnt, 10 to 30rnt, 10 to 20rnt, 20 to 90rnt, 20 to 80rnt, 20 to 70rnt, 20 to 60rnt, 20 to 50rnt, 20 to 40rnt, 30 to 90rnt, 30 to 80rnt, 30 to 70rnt, 30 to 60rnt, or 30 to 50rnt in length.

[0133] In all embodiments, the TREMs described herein have the formula I ZZZ The consensus sequence of R0-R 1- R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x1 -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R72 wherein: ZZZ represents any of 20 amino acids; (ii) Formula I corresponds to all species; (iii) x = 1 to 271 (e.g., x = 1 to 250, x = 1 to 225, x = 1 to 200, x = 1 to 175, x = 1 to 150, x = 1 to 125, x = 1 to 100, x = 1 to 75, x = 1 to 50, x = 1 to 40, x = 1 to 30, x = 1 to 29, x = 1 to 28, x = 1 to 27, x = 1 to 26) , x=1~25, x=1~24, x=1~23, x=1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~ 14, x=1~13, x=1~12, x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~27 1, x=70~271, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x= 1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19 , x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271; (iv) the ASGPR binding moiety is R0-R 1- or (v) the ASGPR binding moiety is attached to a nucleobase within one or more of R2-R3-R4-R5-R6-R7-R8; 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 is attached to one or more of the nucleobases.

[0134] In all embodiments, the TREMs described herein have the formula II ZZZ The consensus sequence of R0-R 1- R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x1 -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67-R 68 -R 69 -R 70 -R 71 -R 72 wherein: ZZZ represents any of 20 amino acids; (ii) Formula II corresponds to mammals; (iii) x = 1 to 271 (e.g., x = 1 to 250, x = 1 to 225, x = 1 to 200, x = 1 to 175, x = 1 to 150, x = 1 to 125, x = 1 to 100, x = 1 to 75, x = 1 to 50, x = 1 to 40, x = 1 to 30, x = 1 to 29, x = 1 to 28, x = 1 to 27, x = 1 to 26) , x=1~25, x=1~24, x=1~23, x=1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~ 14, x=1~13, x=1~12, x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~27 1, x=70~271, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x= 1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19 , x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271; (iv) the ASGPR binding moiety is R0-R 1- or (v) the ASGPR binding moiety is attached to a nucleobase within one or more of R2-R3-R4-R5-R6-R7-R8; 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71-R 72 is attached to one or more of the nucleobases.

[0135] In all embodiments, the TREMs described herein are represented by formula IIII ZZZ The consensus sequence of R0-R 1- R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x1 -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 wherein: ZZZ represents any of 20 amino acids; (ii) Formula III corresponds to human; (iii) x = 1 to 271 (e.g., x = 1 to 250, x = 1 to 225, x = 1 to 200, x = 1 to 175, x = 1 to 150, x = 1 to 125, x = 1 to 100, x = 1 to 75, x = 1 to 50, x = 1 to 40, x = 1 to 30, x = 1 to 29, x = 1 to 28, x = 1 to 27, x = 1 to 26) , x=1~25, x=1~24, x=1~23, x=1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~ 14, x=1~13, x=1~12, x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~27 1, x=70~271, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x= 1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19 , x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271; (iv) the ASGPR binding moiety is R0-R 1- or (v) the ASGPR binding moiety is attached to a nucleobase within one or more of R2-R3-R4-R5-R6-R7-R8; 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 is attached to one or more of the nucleobases.

[0136] [Table 3]

[0137] [Table 4]

[0138] [Table 5]

[0139] [Table 6]

[0140] [Table 7]

[0141] [Table 8]

[0142] [Table 9]

[0143] [Table 10]

[0144] [Table 11]

[0145]

Table 12

[0146]

Table 13

[0147]

Table 14

[0148]

Table 15

[0149] Table 16

[0150]

Table 17

[0151]

Table 18

[0152]

Table 19

[0153] Table 20

[0154] Table 21

[0155] Table 22

[0156] [Table 23]

[0157] [Table 24]

[0158] Asialoglycoprotein receptor binding site The present disclosure features TREMs containing an asialoglycoprotein receptor (ASGPR)-binding moiety. ASGPR is a C-type lectin primarily expressed on the sinusoidal surface of hepatocytes and contains a major (48 kDa, ASGPR-1) and a minor (40 kDa, ASGPR-2) subunit. ASGPR is involved in the binding, internalization, and subsequent clearance of glycoproteins containing N-terminal galactose (Gal) or N-acetylgalactosamine (GalNAc) residues from the circulation, such as antibodies. ASGPR has also been shown to be involved in the clearance of low-density lipoproteins, fibronectin, and certain immune cells, and may be used by certain viruses to enter hepatocytes (see, e.g., Yang J., et al. (2006) J Viral Hepat 13:158-165 and Guy, C.S. et al. (2011) Nat Rev Immunol 8:874-887).

[0159] The ASGPR-binding moiety described herein may refer to a structure comprising (i) an ASGPR carbohydrate and (ii) an ASGPR linker (e.g., a linker connecting the carbohydrate to a TREM). The term "carbohydrate" as used herein refers to a compound comprising one or more monosaccharide moieties comprising at least three carbon atoms (e.g., arranged in a linear, branched, or cyclic configuration) and an oxygen, nitrogen, or sulfur atom, or a fragment or variant of a monosaccharide moiety comprising at least three carbon atoms (e.g., arranged in a linear, branched, or cyclic configuration) and an oxygen, nitrogen, or sulfur atom. Each monosaccharide moiety or fragment or variant thereof may be a tetrose, pentose, hexose, or heptose. Each monosaccharide moiety or fragment or variant thereof may exist as an aldose, ketose, sugar alcohol, or in the L- or D-form, as appropriate. Exemplary monosaccharide moieties may be amino sugars, N-acetylamino sugars, imino sugars, deoxy sugars, or sugar acids. Carbohydrates may include individual monosaccharide moieties, or may further include disaccharides, oligosaccharides (e.g., trisaccharides, tetrasaccharides, pentasaccharides, hexasaccharides, heptasaccharides, octasaccharides), polysaccharides, or combinations thereof.Exemplary carbohydrates include ribose, arabinose, lyxose, xylose, deoxyribose, ribulose, xylulose, glucose, galactose, mannose, gulose, idose, talose, allose, altrose, psicose, fructose, sorbose, tagatose, rhamnose, pneumose, quinovose, fucose, mannoheptulose, sedoheptulose, galactosamine, mannosamine, glucosamine, N-acetylglucosamine, N-acetylgalactosamine, N-acetylmannosamine, glucuronic acid, galacturonic acid, mannuronic acid, guluronic acid, iduronic acid, tagaturonic acid, frucuronic acid, galactosaminuronic acid, mannosaminuronic acid, glucosaminuronic acid, and the like. Examples of sugars that can be used include N-acetylglucosaminuronic acid, N-acetylgalactosaminuronic acid, N-acetylmannosaminuronic acid, maltose, lactose, sucrose, trehalose, gentiobiose, cellobiose, chitobiose, kojibiose, nigerose, sophorose, trehalulose, isomaltose, xylobiose, starch, cellulose, chitin, and dextran.

[0160] A carbohydrate can include one or more monosaccharide moieties linked by glycosidic bonds. In some embodiments, the glycosidic bonds include 1 to 2 glycosidic bonds, 1 to 3 glycosidic bonds, 1 to 4 glycosidic bonds, or 1 to 6 glycosidic bonds. In some embodiments, each glycosidic bond can be in the α or β configuration. In certain embodiments, the one or more monosaccharide moieties are directly linked by glycosidic bonds or separated by a linker.

[0161] In some embodiments, the ASGPR-binding moiety comprises a galactose (Gal), galactosamine (GalNH), or N-acetylgalactosamine (GalNAc) moiety, e.g., Gal, GalNH, or GalNAc, or an analog thereof. In some embodiments, the ASGPR-binding moiety comprises a GalNAc moiety (e.g., GalNAc). In some embodiments, the ASGPR-binding moiety comprises multiple GalNAc moieties (e.g., GalNAc), e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more GalNAc moieties (e.g., GalNAc). In some embodiments, the ASGPR-binding moiety comprises 2 to 20 GalNAc moieties (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 GalNAc moieties). In some embodiments, the ASGPR-binding moiety comprises 2 to 10 GalNAc moieties (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 GalNAc moieties). In some embodiments, the ASGPR-binding moiety comprises 2 to 5 GalNAc moieties (e.g., 2, 3, 4, or 5 GalNAc moieties). In some embodiments, the ASGPR-binding moiety comprises 2 GalNAc moieties. In some embodiments, the ASGPR-binding moiety comprises 3 GalNAc moieties. In some embodiments, the ASGPR-binding moiety comprises 4 GalNAc moieties. In some embodiments, the ASGPR-binding moiety comprises 5 GalNAc moieties.

[0162] In some embodiments, the GalNAc moiety has the structure of formula (I): [ka] or a salt thereof, wherein each of X and Y is independently O, N(R 7 ), or S;R 1 , R 3 , R 4 , and R 5is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, C(O)-alkyl, C(O)-alkenyl, C(O)-alkynyl, C(O)-heteroalkyl, C(O)-haloalkyl, C(O)-aryl, C(O)-heteroaryl, C(O)-cycloalkyl, or C(O)-heterocyclyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 8 optionally substituted with; or R 3 and R 4 together with the oxygen atoms to which they are attached, form one or more R 8 forming a heterocyclyl ring optionally substituted with R 2a is hydrogen or alkyl; R 2b is —C(O)alkyl (e.g., C(O)CH); R 6a and R 6b each of which is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, halo, cyano, nitro, -OR A , aryl, heteroaryl, cycloalkyl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 9 optionally substituted with ;R 7 is hydrogen, alkyl, or C(O)-alkyl; R 8 and R 9 each is independently hydrogen, halo, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, cycloalkyl, or heterocyclyl; R A is hydrogen, or alkyl, alkenyl, or alkynyl, and n is an integer of 0 to 6, where the structure of formula (I) can be attached to a linker or nucleobase within the ASt of the TREM.

[0163] In some embodiments, X is O. In some embodiments, Y is O. In some embodiments, R 1 , R 3 , R 4 , and R 5 Each of R is independently hydrogen or alkyl (e.g., CH). 2a is hydrogen. In some embodiments, R 2b is C(O)CH3. In some embodiments, R 6a and R 6b are hydrogen. In some embodiments, n is 0, 1, 2, or 3. In some embodiments, n is 1, 2, or 3. In some embodiments, n is 1. In some embodiments, the GalNAc moiety is 2a In some embodiments, the GalNAc moiety is attached to a linker or TREM at R 2b In some embodiments, the GalNAc moiety is attached to a linker or TREM at R 3 In some embodiments, the GalNAc moiety is attached to a linker or TREM at R 4 In some embodiments, the GalNAc moiety is attached to a linker or TREM at R 5 In some embodiments, the GalNAc moiety is attached to a linker or TREM at R 6a or R 6b In some embodiments, the GalNAc moiety is attached to a linker or TREM at multiple positions, e.g., R 1 , R 2a , R 2b , R 3 , R 4 , R 5 , R 6a , and R 6b At least two of the amino acid sequences are linked to a linker or a TREM.

[0164] In some embodiments, the GalNAc moiety has the structure of formula (Ia) [ka] or a salt thereof, wherein R 2a is hydrogen or alkyl; R 2b is —C(O)alkyl (e.g., C(O)CH); R 3 , R 4 , and R 5 is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, C(O)-alkyl, C(O)-alkenyl, C(O)-alkynyl, C(O)-heteroalkyl, C(O)-haloalkyl, C(O)-aryl, C(O)-heteroaryl, C(O)-cycloalkyl, or C(O)-heterocyclyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 8 optionally substituted with; or R 3 and R 4 together with the oxygen atoms to which they are attached, form one or more R 8 forming a heterocyclyl ring optionally substituted with R 8 is hydrogen, halo, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, cycloalkyl, or heterocyclyl, wherein [ka] represents a bond in any configuration, [ka] represents the point of attachment to the TREM, for example, a linker, a nucleobase, an internucleotide bond, or a terminus within the TREM sequence.

[0165] In some embodiments, R 3 , R 4 , and R 5 Each of R is independently hydrogen or alkyl (e.g., CH). 2a is hydrogen. In some embodiments, R2b is C(O)CH3.

[0166] In some embodiments, the GalNAc moiety has the structure of formula (II): [ka] or a salt thereof, wherein X is O, N(R 7 ), or S; each of W or Y is independently O or C(R 10a )(R 10b ) where one of W and Y is O; R 1 , R 3 , R 4 , and R 5 is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, C(O)-alkyl, C(O)-alkenyl, C(O)-alkynyl, C(O)-heteroalkyl, C(O)-haloalkyl, C(O)-aryl, C(O)-heteroaryl, C(O)-cycloalkyl, or C(O)-heterocyclyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 8 optionally substituted with; or R 3 and R 4 together with the oxygen atoms to which they are attached, form one or more R 8 forming a heterocyclyl ring optionally substituted with R 2a is hydrogen or alkyl; R 2b is —C(O)alkyl (e.g., C(O)CH); R 6a and R 6b each of which is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, halo, cyano, nitro, -OR A , aryl, heteroaryl, cycloalkyl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R9 optionally substituted with ;R 7 is hydrogen, alkyl, or C(O)-alkyl; R 8 and R 9 each is independently hydrogen, halo, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, cycloalkyl, or heterocyclyl; R 10a and R 10b each of R is independently hydrogen, heteroalkyl, haloalkyl, or halo; A is hydrogen, or alkyl, alkenyl, alkynyl, where the structure of formula (I) can be attached to a TREM, for example, to a linker, nucleobase, internucleotide bond, or terminus within a TREM sequence.

[0167] In some embodiments, the GalNAc moiety has the structure of formula (II-a): [ka] or a salt thereof, wherein X is O, N(R 7 ), or S;R 1 , R 3 , R 4 , and R 5 is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, C(O)-alkyl, C(O)-alkenyl, C(O)-alkynyl, C(O)-heteroalkyl, C(O)-haloalkyl, C(O)-aryl, C(O)-heteroaryl, C(O)-cycloalkyl, or C(O)-heterocyclyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 8 optionally substituted with; or R 3 and R 4 together with the oxygen atoms to which they are attached, form one or more R 8 forming a heterocyclyl ring optionally substituted with R 2a is hydrogen or alkyl; R 2bis —C(O)alkyl (e.g., C(O)CH); R 6a and R 6b each of which is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, halo, cyano, nitro, -OR A , aryl, heteroaryl, cycloalkyl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 9 optionally substituted with ;R 7 is hydrogen, alkyl, or C(O)-alkyl; R 8 and R 9 each is independently hydrogen, halo, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, cycloalkyl, or heterocyclyl; R A is hydrogen, or alkyl, alkenyl, alkynyl, where the structure of formula (I) can be attached to a TREM, for example, to a linker, nucleobase, internucleotide bond, or terminus within a TREM sequence.

[0168] In some embodiments, the GalNAc moiety has the structure of formula (II-b): [ka] or a salt thereof, wherein X is O, N(R 7 ), or S;R 1 , R 3 , R 4 , and R 5is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, C(O)-alkyl, C(O)-alkenyl, C(O)-alkynyl, C(O)-heteroalkyl, C(O)-haloalkyl, C(O)-aryl, C(O)-heteroaryl, C(O)-cycloalkyl, or C(O)-heterocyclyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 8 optionally substituted with; or R 3 and R 4 together with the oxygen atoms to which they are attached, form one or more R 8 forming a heterocyclyl ring optionally substituted with R 2a is hydrogen or alkyl; R 2b is —C(O)alkyl (e.g., C(O)CH); R 6a and R 6b each of which is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, halo, cyano, nitro, -OR A , aryl, heteroaryl, cycloalkyl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from the group consisting of one or more R 9 optionally substituted with ;R 7 is hydrogen, alkyl, or C(O)-alkyl; R 8 and R 9 each is independently hydrogen, halo, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, cycloalkyl, or heterocyclyl; R A is hydrogen, or alkyl, alkenyl, alkynyl, where the structure of formula (I) can be attached to a TREM, for example, to a linker, nucleobase, internucleotide bond, or terminus within a TREM sequence.

[0169] In some embodiments, the ASGPR binding moiety has the structure of formula (III): [ka] or a salt thereof, wherein R 1 , R 2a , R 2b , R 3 , R 4 , R 5 , R 6a , and R 6b and subvariables thereof are as defined for formula (I), L is a linker, and n is an integer from 1 to 100, wherein: [ka] represents the point of attachment to the branch point, a further linker, or a TREM, for example a linker, nucleobase, internucleotide bond, or terminus within the TREM sequence.

[0170] In some embodiments, X is O. In some embodiments, R 1 , R 3 , R 4 , and R 5 Each of R is independently hydrogen or alkyl (e.g., CH). 2a is hydrogen. In some embodiments, R 2b is C(O)CH3. In some embodiments, R 6a and R 6b are each hydrogen. In some embodiments, n is an integer from 1 to 50. In some embodiments, n is an integer from 1 to 25. In some embodiments, n is an integer from 1 to 10. In some embodiments, n is an integer from 1 to 5. In some embodiments, n is 1, 2, 3, 4, or 5. In some embodiments, n is 1.

[0171] In certain embodiments, L comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. In certain embodiments, L comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. In certain embodiments, L is cleavable or non-cleavable.

[0172] The term "linker," as used herein, refers to an organic moiety that connects two or more portions of a compound, for example, via a covalent bond. A linker can be linear or branched. In some embodiments, a linker comprises a heteroatom, such as a nitrogen, sulfur, oxygen, phosphorus, silicon, or boron atom. In some embodiments, a linker comprises a cyclic group (e.g., an aryl, heteroaryl, cycloalkyl, or heterocyclyl group). In some embodiments, a linker comprises a functional group, such as an amide, ketone, ester, ether, thioester, thioether, thiol, hydroxyl, amine, cyano, nitro, azide, triazole, pyrroline, p-nitrophenyl, alkene, or alkyne group. Any atom in a linker can be substituted or unsubstituted.In some embodiments, the linker is an arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkynyl and alkylaryl, alkenylaryl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl, alkynylheteroarylalkynyl, alkylheterocyclylalkyl, alkylheterocyclylalkenyl, alkylheterocyclyl(alkynyl)alkynyl, alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl, alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl, or alkynylheteroaryl(alkynyl) groups. In some embodiments, the linker comprises a polyethylene glycol group (e.g., PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG10, PEG12, PEG14, PEG16, PEG18, PEG20, PEG24, PEG28, PEG32, PEG100, PEG200, PEG250, PEG500, PEG600, PEG700, PEG750, PEG800, PEG900, PEG1000, PEG2000, or PEG3000). In some embodiments, L comprises a PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 group.In some embodiments, L comprises multiple PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups (e.g., 2, 3, 4, or 5 PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups). In some embodiments, L comprises a PEG2 group. In some embodiments, L comprises multiple PEG2 groups. In some embodiments, L comprises a PEG3 group. In some embodiments, L comprises multiple PEG3 groups. In some embodiments, L comprises a PEG4 group. In some embodiments, L comprises multiple PEG4 groups.

[0173] In some embodiments, the linker comprises 1 to 1000 atoms (e.g., 1 to 750 atoms, 1 to 500 atoms, 1 to 250 atoms, 1 and 100 atoms, 1 to 75 atoms, 1 to 50 atoms, 1 to 25 atoms, and 1 to 10 atoms). In some embodiments, the linker comprises 1 to 100 atoms. In some embodiments, the linker comprises 1 to 50 atoms. In some embodiments, the linker comprises 1 to 25 atoms.

[0174] In some embodiments, the linker is linear and comprises 1 to 1000 atoms (e.g., 1 to 750 atoms, 1 to 500 atoms, 1 to 250 atoms, 1 to 100 atoms, 1 to 75 atoms, 1 to 50 atoms, 1 to 25 atoms, and 1 to 10 atoms). In some embodiments, the linker is linear and comprises 1 to 100 atoms. In some embodiments, the linker is linear and comprises 1 to 50 atoms. In some embodiments, the linker is linear and comprises 1 to 25 atoms.

[0175] In some embodiments, the linker is branched and each branch comprises 1 to 1000 atoms (e.g., 1 to 750 atoms, 1 to 500 atoms, 1 to 250 atoms, 1 to 100 atoms, 1 to 75 atoms, 1 to 50 atoms, 1 to 25 atoms, and 1 to 10 atoms). In some embodiments, the linker is branched and each branch comprises 1 to 100 atoms. In some embodiments, the linker is branched and each branch comprises 1 to 50 atoms. In some embodiments, the linker is branched and each branch comprises 1 to 25 atoms.

[0176] In some embodiments, the ASGPR binding moiety has the structure of formula (III-a): [ka] or a salt thereof, wherein R 1 , R 2a , R 2b , R 3 , R 4 , R 5 , R 6a , and R 6b and each of the subvariables thereof are as defined for formula (I), and L 1 and L 2 each independently represents a linker, each of m and n independently represents an integer from 1 to 100, and M represents a linker, [ka] represents the point of attachment to the branch point, a further linker, or a TREM, for example a linker, nucleobase, internucleotide bond, or terminus within the TREM sequence.

[0177] In some embodiments, X is O (e.g., X in each of A and B is O). In some embodiments, R 1 , R 3 , R 4 , and R 5is independently hydrogen or alkyl (e.g., CH) (e.g., R in each of A and B). 1 , R 3 , R 4 , and R 5 are independently hydrogen or alkyl. In some embodiments, R 2a is hydrogen (e.g., R 2a is hydrogen). In some embodiments, R 2b is C(O)CH3 (for example, R 2b is C(O)CH. In some embodiments, R 6a and R 6b each of A and B is hydrogen (e.g., R 6a and R 6b is hydrogen). In some embodiments, each of m and n is independently an integer from 1 to 50. In some embodiments, each of m and n is independently an integer from 1 to 25. In some embodiments, each of m and n is independently an integer from 1 to 10. In some embodiments, each of m and n is independently an integer from 1 to 5. In some embodiments, each of m and n is independently 1, 2, 3, 4, or 5. In some embodiments, each of m and n is independently 1.

[0178] In one embodiment, L 1 and L 2 Each of L independently comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. 1 and L 2 Each of L independently comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. 1 and L 2 Each of L is independently cleavable or non-cleavable. 1 and L 2each independently comprises a polyethylene glycol group (e.g., PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG10, PEG12, PEG14, PEG16, PEG18, PEG20, PEG24, PEG28, PEG32, PEG100, PEG200, PEG250, PEG500, PEG600, PEG700, PEG750, PEG800, PEG900, PEG1000, PEG2000, or PEG3000). 1 and L 2 Each of L independently comprises a PEG, PEG, PEG, PEG, or PEG group. 1 and L 2 each independently comprises multiple PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups (e.g., 2, 3, 4, or 5 PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups). 1 and L 2 Each of L independently comprises a PEG group. 1 and L 2 Each of L independently contains multiple PEG groups. 1 and L 2 Each of L independently comprises a PEG group. 1 and L 2 Each of L independently contains multiple PEG groups. 1 and L 2 Each of L independently comprises a PEG group. 1 and L 2 each independently contains multiple PEG4 groups.

[0179] In some embodiments, M comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. In certain embodiments, M comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. In certain embodiments, M is cleavable or non-cleavable.

[0180] In some embodiments, the ASGPR binding moiety has the structure of formula (III-b): [ka] or a salt thereof, wherein R 1 , R 2a , R 2b , R 3 , R 4 , R 5 , R 6a , and R 6b and each of the subvariables thereof are as defined for formula (I), and L 1 , L 2 , and L 3 each of m, n, and o is independently an integer from 1 to 100, and M is a linker, wherein: [ka] represents the point of attachment to the branch point, a further linker, or a TREM, for example a linker, nucleobase, internucleotide bond, or terminus within the TREM sequence.

[0181] In some embodiments, X is O (e.g., X in each of A, B, and C is O). In some embodiments, R 1 , R 3 , R 4 , and R 5 is independently hydrogen or alkyl (e.g., CH) (e.g., R in each of A, B, and C). 1 , R 3 , R 4 , and R 5are independently hydrogen or alkyl. In some embodiments, R 2a is hydrogen (e.g., R 2a is hydrogen). In some embodiments, R 2b is C(O)CH3 (e.g., R 2b is C(O)CH. In some embodiments, R 6a and R 6b each of A, B, and C is hydrogen (e.g., R 6a and R 6b is hydrogen). In some embodiments, each of m, n, and o is independently an integer from 1 to 50. In some embodiments, each of m, n, and o is independently an integer from 1 to 25. In some embodiments, each of m, n, and o is independently an integer from 1 to 10. In some embodiments, each of m, n, and o is independently an integer from 1 to 5. In some embodiments, each of m, n, and o is independently 1, 2, 3, 4, or 5. In some embodiments, each of m, n, and o is independently 1.

[0182] In one embodiment, L 1 , L 2 , and L 3 Each of L independently comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. 1 , L 2 , and L 3 Each of L independently comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. 1 , L 2 , and L 3 Each of L is independently cleavable or non-cleavable. 1 and L 2 Each of L is independently cleavable or non-cleavable. 1 , L2 , and L 3 each independently comprises a polyethylene glycol group (e.g., PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG10, PEG12, PEG14, PEG16, PEG18, PEG20, PEG24, PEG28, PEG32, PEG100, PEG200, PEG250, PEG500, PEG600, PEG700, PEG750, PEG800, PEG900, PEG1000, PEG2000, or PEG3000). 1 , L 2 , and L 3 Each of L independently comprises a PEG, PEG, PEG, PEG, or PEG group. 1 , L 2 , and L 3 each independently comprises multiple PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups (e.g., 2, 3, 4, or 5 PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups). 1 , L 2 , and L 3 Each of L independently comprises a PEG group. 1 , L 2 , and L 3 Each of L independently contains multiple PEG groups. 1 , L 2 , and L 3 Each of L independently comprises a PEG group. 1 , L 2 , and L 3 Each of L independently contains multiple PEG groups. 1 , L 2 , and L 3 Each of L independently comprises a PEG group. 1 , L 2 , and L 3 each independently contains multiple PEG4 groups.

[0183] In some embodiments, M comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. In certain embodiments, M comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. In certain embodiments, M is cleavable or non-cleavable.

[0184] In some embodiments, the ASGPR binding moiety has the structure of formula (III-c): [ka] or a salt thereof, wherein R 2a , R 2b , R 3 , R 4 , R 5 and subvariables thereof are as defined for formula (I), and L 1 , L 2 , and L 3 each is independently a linker and M is a linker, wherein [ka] represents the point of attachment to the branch point, a further linker, or a TREM, for example a linker, nucleobase, internucleotide bond, or terminus within the TREM sequence.

[0185] In some embodiments, R 3 , R 4 , and R 5 Each of R is independently hydrogen or alkyl (e.g., CH). 2a is hydrogen. In some embodiments, R 2b is C(O)CH3.

[0186] In one embodiment, L 1 , L 2 , and L 3Each of L independently comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. 1 , L 2 , and L 3 Each of L independently comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. 1 , L 2 , and L 3 Each of L is independently cleavable or non-cleavable. 1 and L 2 Each of L is independently cleavable or non-cleavable. 1 , L 2 , and L 3 each independently comprises a polyethylene glycol group (e.g., PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG10, PEG12, PEG14, PEG16, PEG18, PEG20, PEG24, PEG28, PEG32, PEG100, PEG200, PEG250, PEG500, PEG600, PEG700, PEG750, PEG800, PEG900, PEG1000, PEG2000, or PEG3000). 1 , L 2 , and L 3 Each of L independently comprises a PEG, PEG, PEG, PEG, or PEG group. 1 , L 2 , and L 3 each independently comprises multiple PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups (e.g., 2, 3, 4, or 5 PEG1, PEG2, PEG3, PEG4, PEG5, or PEG6 groups). 1 , L 2 , and L 3 Each of L independently comprises a PEG group. 1 , L 2 , and L3 Each of L independently contains multiple PEG groups. 1 , L 2 , and L 3 Each of L independently comprises a PEG group. 1 , L 2 , and L 3 Each of L independently contains multiple PEG groups. 1 , L 2 , and L 3 Each of L independently comprises a PEG group. 1 , L 2 , and L 3 each independently contains multiple PEG4 groups.

[0187] In some embodiments, M comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group. In certain embodiments, M comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group. In certain embodiments, M is cleavable or non-cleavable.

[0188] In some embodiments, the ASGPR binding moiety is: [ka] [ka] [ka] [ka] [ka] [ka] The compound includes a compound selected from:

[0189] In some embodiments, the ASGPR binding moiety is compound (Xi). In some embodiments, the ASGPR binding moiety is compound (X-ii). In some embodiments, the ASGPR binding moiety is compound (X-iii). In some embodiments, the ASGPR binding moiety is compound (X-iv). In some embodiments, the ASGPR binding moiety is compound (Xv). In some embodiments, the ASGPR binding moiety is compound (X-vi). In some embodiments, the ASGPR binding moiety is compound (X-vii). In some embodiments, the ASGPR binding moiety is compound (X-viii). In some embodiments, the ASGPR binding moiety is compound (X-ix). In some embodiments, the ASGPR binding moiety is compound (Xx). In some embodiments, the ASGPR binding moiety is compound (X-xi). In some embodiments, the ASGPR binding moiety is compound (X-xii). In some embodiments, the ASGPR binding moiety is compound (X-xiii). In some embodiments, the ASGPR binding moiety is compound (X-xiv). In some embodiments, the ASGPR binding moiety is compound (X-xv). In some embodiments, the ASGPR binding moiety is compound (X-xvi). In some embodiments, the ASGPR binding moiety is compound (X-xvii). In some embodiments, the ASGPR binding moiety is compound (X-xviii). In some embodiments, the ASGPR binding moiety is compound (X-xix). In some embodiments, the ASGPR binding moiety is compound (X-xx). In some embodiments, the ASGPR binding moiety is compound (X-xxi). In some embodiments, the ASGPR binding moiety is compound (X-xxii). In some embodiments, the ASGPR binding moiety is compound (X-xxiii). In some embodiments, the ASGPR binding moiety is a compound selected from compounds (Xi), (X-xxii), and (X-xxii).

[0190] In some embodiments, the ASGPR binding moiety comprises a linker comprising a cyclic moiety, such as a pyrroline ring. In certain embodiments, the ASGPR binding moiety has the structure of formula (CII): [ka] or a salt thereof, wherein E is absent or C(O), C(O)O, C(O)NH, C(S), C(S)NH, SO, SO, or SONH; R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 Each occurrence of is independently H, -CH2OR a OR b and;R a and R b each independently at each occurrence is selected from hydrogen, a hydroxyl protecting group, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted aralkyl, optionally substituted alkenyl, optionally substituted heteroaryl, polyethylene glycol (PEG), phosphate, diphosphate, triphosphate, phosphonate, phosphonothioate, phosphonodithioate, phosphorothioate, phosphorothiolate, phosphorodithioate, phosphorothiolothionate, phosphodiester, phosphotriester, activated phosphate group, activated phosphite group, phosphoramidite, solid support, -P(Z 1 )(Z 2 )-O-nucleosides, -P(Z 1 )(Z 2 )-O-oligonucleotide, -P(Z 1 )(O-Linker-R L )-O-nucleoside, or -P(Z 1 )(O-Linker-R L )-O-oligonucleotide; R 30 Each occurrence of -linker-R L or R 31 and;R Lis hydrogen or a ligand; R 31 -C(O)CH(N(R 32 )2)(CH2) h N(R 32 )2;R 32 independently, for each occurrence, H,-R L , -linker-R L or R 31 and Z 1 is, independently at each occurrence, O or S; Z 2 is independently at each occurrence O, S, N(alkyl) or optionally substituted alkyl; and h is independently at each occurrence 1 to 20.

[0191] In some embodiments, the compound of formula (CII) is [ka] [ka] is selected from.

[0192] In some embodiments, the ASGPR binding moiety is a compound or substructure disclosed in US Pat. No. 8,106,022, which is incorporated herein by reference in its entirety.

[0193] In some embodiments, the ASGPR binding moiety is compound (CII-i). In some embodiments, the ASGPR binding moiety is compound (CII-ii). In some embodiments, the ASGPR binding moiety is compound (CII-iii). In some embodiments, the ASGPR binding moiety is compound (CII-iv). In some embodiments, the ASGPR binding moiety is compound (CII-v). In some embodiments, the ASGPR binding moiety is compound (CII-vi).

[0194] In some embodiments, the ASGPR binding moiety is a compound of formula (C-1), (C-2), (C-3), or (C4): [ka] or a pharmaceutically acceptable salt thereof, wherein: n is 1, 2, or 3; W is absent or a peptide; and L is -(TQTQ) m -, where each T is independently absent or (C1 to C 10 ) alkylene, (C2-C 10 ) alkenylene, or (C2-C 10 ) alkynylene, wherein one or more carbon groups of said T are each independently selected from -O-, -S-, and -N(R 4 )-, where the heteroatom groups are separated by at least two carbon atoms, and where alkylene, alkenylene, and alkynylene can each be independently substituted with one or more halo atoms; each Q is independently absent or selected from C(O), C(O)-NR 4 , N.R. 4 -C(O), OC(O)-NR 4 , N.R. 4 -C(O)-O, -CH2-, heteroaryl, or O, S, SS, S(O), S(O)2, and NR 4 and wherein at least two carbon atoms are selected from any other heteroatomic group selected from the heteroatomic groups O, S, SS, S(O), S(O)2 and NR 4 Separately; each R 4 Independently, -H, -(C1~C 20 ) alkyl, or (C3-C8) cycloalkyl, wherein 1 to 6 -CH2- groups of the alkyl or cycloalkyl separated by at least 2 carbon atoms are -O-, -S-, or -N(R 4 )-, and each -CH3- of the alkyl may be independently substituted with -N(R 4 ) 2、 -OR 4 , and -S(R 4wherein the heteroatom groups are separated by at least two carbon atoms; wherein alkyl and cycloalkyl can be substituted with halo atoms; and m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40.

[0195] In some embodiments, the ASGPR binding moiety is compound (C-1). In some embodiments, the ASGPR binding moiety is compound (C-2). In some embodiments, the ASGPR binding moiety is compound (C-3). In some embodiments, the ASGPR binding moiety is compound (C-4).

[0196] In some embodiments, the compound of formula (C-1), (C-2), (C-3), or (C4) is [ka] (wherein n' is 1 or 2) or a pharmaceutically acceptable salt thereof.

[0197] In some embodiments, the ASGPR binding moiety is a compound of formula (E): [ka] or a pharmaceutically acceptable salt thereof, wherein: n is i, 2, or 3; W is absent or a peptide; and L is -(TQTQ) m -, where each T is independently absent or (C1 to C 10 ) alkylene, (C2-C 10 ) alkenylene, or (C2-C 10 ) alkynylene, wherein one or more carbon groups of said T are each independently selected from -O-, -S-, and -N(R 4)-, wherein the heteroatom groups are separated by at least two carbon atoms, and wherein said alkylene, alkenylene, alkynylene may each be independently substituted with one or more halo atoms; each Q is independently absent or selected from C(O), C(O)-R 4 , R 4 -C(O), OC(O)-R 4 , R 4 -C(O)-O, -CH2-, heteroaryl, or O, S, SS, S(O), S(O)2, and NR 4 and wherein at least two carbon atoms are selected from any other heteroatomic group selected from the heteroatomic groups O, S, SS, S(O), S(O)2 and NR 4 Separately; each R 4 Independently, -H, -(C1~C 20 ) alkyl, -(C1-C 20 ) alkenyl, -(C2-C 20 )alkynyl, or (C3-C6)cycloalkyl, wherein 1 to 6 -CH2- groups of the alkyl or cycloalkyl separated by at least 2 carbon atoms are -O-, -S-, or -N(R 4 )-, and the -CH3 of the alkyl may be substituted with -N(R 4 )2, -OR 4 , and -S(R 4 wherein the heteroatom groups are separated by at least two carbon atoms; and wherein alkyl, alkenyl, alkynyl, and cycloalkyl can be substituted with halo atoms; and each m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40.

[0198] In some embodiments, the compound of formula (E) is [ka] or a pharmaceutically acceptable salt thereof, and Y is as defined in formula (E).

[0199] In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0200] In some embodiments of the compound of Formula (E), the compound is [ka] or a pharmaceutically acceptable salt thereof.

[0201] In some embodiments, the ASGPR binding moiety is a compound or substructure disclosed in WO 2017 / 083368, which is incorporated herein by reference in its entirety.

[0202] In other embodiments, the ASGPR binding moiety is: [ka] [ka] wherein one of X or Y is a branch point, a linker, or a TREM, e.g., a linker, nucleobase, internucleotide bond, or terminus within a TREM sequence, and the other of X and Y is hydrogen.

[0203] In certain embodiments, the ASGPR binding moiety has the structure of formula (XII-a): [ka] In certain embodiments, the ASGPR binding moiety is a compound or substructure disclosed in Nucleic Acids (2016) 5:e317 or WO 2015 / 042447, each of which is incorporated by reference in its entirety.

[0204] In some embodiments, the ASGPR binding moiety has the structure of formula (Va): [ka] wherein n is an integer from 1 to 20. In some embodiments, the compound of Formula (Va) comprises: [ka] wherein Z is a linker or nucleobase in an oligomeric compound, such as ASt of a TREM.

[0205] In another embodiment, the ASGPR binding moiety has the structure of formula (Vb): [ka] wherein A is O or S, A' is O, S, or NH, and Z is an oligomeric compound, e.g., a linker or a TREM, e.g., a linker, nucleobase, internucleotide linkage, or terminus within a TREM sequence.

[0206] In some embodiments, the ASGPR binding moiety is: [ka] Includes:

[0207] In some embodiments, the ASGPR binding moiety is: [ka] is selected from.

[0208] In certain embodiments, the ASGPR binding moiety is a compound or substructure disclosed in WO 2017 / 156012, which is incorporated herein by reference in its entirety.

[0209] In some embodiments, hydroxyl groups in the ASGPR binding moiety are protected, for example, with acetyl or acetonide moieties. In some embodiments, hydroxyl groups in the ASGPR binding moiety are protected with acetyl groups. In some embodiments, hydroxyl groups in the ASGPR binding moiety are protected with acetonide groups. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more hydroxyl groups in the ASGPR binding moiety can be protected, for example, with acetyl or acetonide groups. In some embodiments, all of the hydroxyl groups in the ASGPR binding moiety are protected.

[0210] Exemplary TREMs comprising an ASGPR-binding moiety can have a binding affinity for ASGPR of 0.01 nM to 100 mM. In some embodiments, a TREM comprising an ASGPR-binding moiety has a binding affinity of less than 10 mM, e.g., 7.5 mM, 5 mM, 2.5 mM, 1 mM, 0.75 mM, 0.5 mM, 0.25 mM, 0.1 mM, 75 nM, 50 nM, 25 nM, 10 nM, 5 nM, or less.

[0211] The exemplary TREM that comprises ASGPR binding moiety can be taken up into cells, for example, into hepatocytes.In some embodiments, the TREM that comprises ASGPR binding moiety has increased cellular uptake compared with the TREM that does not comprise ASGPR binding moiety.For example, the TREM that comprises ASGPR binding moiety can be taken up into cells at 1.1, 1.2, 1.3, 1.4, 1.5, 1.75, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100 times or more than the TREM that does not comprise ASGPR binding moiety.

[0212] Further exemplary ASGPR moieties are described in further detail in U.S. Pat. Nos. 8,828,956; 9,867,882; 10,450,568; 10,808,246; U.S. Patent Application Publication Nos. 2015 / 0246133; 2015 / 0203843; and 2012 / 0095200; and PCT Publication Nos. WO 2013 / 166155, WO 2012 / 030683, and WO 2013 / 166121 (each of which is incorporated by reference herein in its entirety).

[0213] ASGPR linker The ASGPR-binding moiety comprises at least one linker that connects the carbohydrate to the TREM. In some embodiments, the TREM is linked to one or more carbohydrates (e.g., the GalNAc moiety of Formula (I)) via a linker described herein. The linker can be monovalent or multivalent, e.g., bivalent, trivalent, tetravalent, or pentavalent. In some embodiments, the linker is [ka] wherein q2A, q2B, q3A, q3B, q4A, q4B, q5A, q5B, and q5C independently represent, at each occurrence, 0 to 20, and wherein the repeating units may be the same or different; P 2A , P 2B , P 3A , P 3B , P 4A , P 4B , P 5A , P 5B , P 5C , T 2A , T 2B , T 3A , T 3B , T 4A , T 4B , T 4A , T 5B , T 5Cis each independently at each occurrence absent, CO, NH, O, S, OC(O), NHC(O), CH, CHNH, or CHO; Q 2A , Q 2B , Q 3A , Q 3B , Q 4A , Q 4B , Q 5A , Q 5B , Q 5C is independently, each occurrence, absent, alkylene, or substituted alkylene, where one or more methylenes are selected from O, S, S(O), SO, N(R N ), C(R')=C(R''), C≡C, or C(O); R 2A , R 2B , R 3A , R 3B , R 4A , R 4B , R 5A , R 5B , R 5C Each occurrence independently represents absent, NH, O, S, CH2, C(O)O, C(O)NH, NHCH(R a )C(O), -C(O)-CH(R a )-NH-, CO, CH=NO, [ka] or heterocyclyl; L 2A , L 2B , L 3A , L 3B , L 4A , L 4B , L 5A , L 5B and L 5C represents a ligand; i.e., each independently at each occurrence is a monosaccharide (e.g., GalNAc), disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide; R a is H or an amino acid side chain.

[0214] In some embodiments, the linker is [ka] wherein L 5A , L 5B and L 5C represents a monosaccharide, such as, for example, a GalNAc derivative, as described herein.

[0215] A cleavable tether is one that is sufficiently stable outside a cell, but after entry into a target cell, is cleaved to release the two moieties held together by the linker. In preferred embodiments, the cleavable tether is cleaved at least about 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold or more, or at least about 100-fold faster inside the target cell or under first reference conditions (which may, for example, be selected to mimic or represent intracellular conditions) than in the subject's blood or under second reference conditions (which may, for example, be selected to mimic or represent conditions found in blood or serum).

[0216] Cleavable linking groups are sensitive to cleaving agents, such as pH, redox potential, or the presence of degradative molecules. Generally, cleaving agents are found more abundantly or at higher levels or activity inside cells than in serum or blood. Examples of such degradative agents include, for example, oxidizing or reducing enzymes or reducing agents, such as mercaptans, present in cells that can degrade redox-cleavable linking groups by reduction; esterases; endosomes or agents that can create an acidic environment, e.g., those that result in a pH of 5 or less; redox agents that are selected for specific substrates or have no substrate specificity, including enzymes that can hydrolyze or degrade acid-cleavable linking groups by acting as general acids, peptidases (which may be substrate-specific), and phosphatases.

[0217] Cleavable linking groups, such as disulfide bonds, can be pH-sensitive. While the pH of human serum is 7.4, the average intracellular pH is slightly lower, ranging from about 7.1 to 7.3. Endosomes have a more acidic pH, ranging from 5.5 to 6.0, and lysosomes have an even more acidic pH of about 5.0. Some linkers will have cleavable linking groups that are cleaved at a preferred pH, thereby releasing the cationic lipid from the ligand inside the cell or into a desired compartment of the cell.

[0218] The linker may contain a cleavable linking group that can be cleaved by a specific enzyme. The type of cleavable linking group incorporated into the linker may depend on the target cell. For example, a liver targeting ligand may be linked to a cationic lipid via a linker containing an ester group. Liver cells are rich in esterases, and therefore the linker will be cleaved more efficiently in liver cells than in cell types that are not rich in esterases. Other cell types that are rich in esterases include lung, renal cortex, and testicular cells. Linkers containing peptide bonds may be used when targeting peptidase-rich cell types, such as hepatocytes and synovial cells.

[0219] In general, the suitability of a candidate cleavable linking group can be evaluated by testing the ability (or conditions) of a degradation agent to cleave the candidate linking group. It may also be desirable to test the candidate cleavable linking group for its ability to resist cleavage in blood or when in contact with other non-target tissues. Thus, the relative susceptibility to cleavage can be determined between first and second conditions, where the first condition is selected to indicate cleavage within target cells and the second condition is selected to indicate cleavage in other tissues or body fluids, such as blood or serum. Evaluation can be performed in a cell-free system, in cells, in cell culture, in organ or tissue culture, or in whole animals. It may be useful to perform initial evaluations in cell-free or culture conditions and confirm with further evaluations in whole animals. In preferred embodiments, useful candidate compounds are cleaved at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times faster inside cells (or under in vitro conditions selected to mimic intracellular conditions) compared to in blood or serum (or under in vitro conditions selected to mimic extracellular conditions).

[0220] In some embodiments, the cleavable linker is a redox-cleavable linker that is cleaved after reduction or oxidation. An example of a reductively cleavable linker is a disulfide linker (-SS-). To determine whether a candidate cleavable linker is a suitable "reductively cleavable linker," or, for example, whether it is suitable for use with a particular TREM moiety and a particular targeting agent, one can rely on the methods described herein. For example, candidates can be evaluated by incubation with dithiothreitol (DTT) or other reducing agents using reagents known in the art that mimic the cleavage rate that would be observed in cells, e.g., target cells. Candidates can also be evaluated under conditions selected to mimic blood or serum conditions. In some embodiments, the candidate compound is cleaved by about 10% or less in blood. In other embodiments, useful candidate compounds are degraded at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times faster inside cells (or under in vitro conditions selected to mimic intracellular conditions) compared to blood (or under in vitro conditions selected to mimic extracellular conditions). The cleavage rate of the candidate compound can be determined using standard enzyme kinetic assays under conditions selected to mimic intracellular medium and compared to conditions selected to mimic extracellular medium.

[0221] In another embodiment, the cleavable linker comprises a phosphate-based cleavable linker that is cleaved by an agent that degrades or hydrolyzes the phosphate group. An example of an agent that cleaves phosphate groups within a cell is an enzyme such as an intracellular phosphatase. Examples of phosphate-based linking groups are -OP(O)(ORk)-O-, -OP(S)(ORk)-O-, -OP(S)(SRk)-O-, -SP(O)(ORk)-O-, -OP(O)(ORk)-S-, -SP(O)(ORk)-S-, -OP(S)(ORk)-S-, -SP(S)(ORk)-O-, -OP(O)(Rk)-O-, -OP(S)(Rk)-O-, -SP(O)(Rk)-O-, -SP(S)(Rk)-O-, -SP(O)(Rk)-S-, -OP(S)(Rk)-S-. Preferred embodiments are -OP(O)(OH)-O-, -OP(S)(OH)-O-, -OP(S)(SH)-O-, -SP(O)(OH)-O-, -OP(O)(OH)-S-, -SP(O)(OH)-S-, -OP(S)(OH)-S-, -SP(S)(OH)-O-, -OP(O)(H)-O-, -OP(S)(H)-O-, -SP(O)(H)-O, -SP(S)(H)-O-, -SP(O)(H)-S-, -OP(S)(H)-S-. A preferred embodiment is -OP(O)(OH)-O-. These candidates can be evaluated using methods similar to those described above.

[0222] In another embodiment, the cleavable linker comprises an acid-cleavable linking group. An acid-cleavable linking group is a linking group that is cleaved under acidic conditions. In a preferred embodiment, the acid-cleavable linking group is cleaved in an acidic environment having a pH of about 6.5 or less (e.g., about 6.0, 5.75, 5.5, 5.25, 5.0, or less) or by an agent, such as an enzyme, that can act as a general acid. Within a cell, certain low-pH organelles, such as endosomes and lysosomes, may provide a cleavage environment for the acid-cleavable linking group. Examples of acid-cleavable linking groups include, but are not limited to, hydrazones, esters, and esters of amino acids. Acid-cleavable groups may have the general formula -C=NN-, C(O)O, or -OC(O). A preferred embodiment is when the carbon bonded to the oxygen of the ester (alkoxy group) is an aryl group, a substituted alkyl group, or a tertiary alkyl group, such as dimethylpentyl or t-butyl. These candidates can be evaluated using methods similar to those described above.

[0223] In another embodiment, the cleavable linker comprises an ester-based cleavable linker. Ester-based cleavable linkers are cleaved by enzymes such as intracellular esterases and amidases. Examples of ester-based cleavable linkers include, but are not limited to, esters of alkylene, alkenylene, and alkynylene groups. Ester cleavable linkers have the general formula -C(O)O- or -OC(O)-. These candidates can be evaluated using methods similar to those described above.

[0224] In yet another embodiment, the cleavable linker comprises a peptide-based cleavable tether. Peptide-based cleavable tethers are cleaved by enzymes such as intracellular peptidases and proteases. Peptide-based cleavable tethers are peptide bonds formed between amino acids to give rise to oligopeptides (e.g., dipeptides, tripeptides, etc.) and polypeptides. Peptide-based cleavable tethers do not include amide groups (—C(O)NH—). Amide groups can be formed between any alkylene, alkenylene, or alkynelene. A peptide bond is a special type of amide bond formed between amino acids to give rise to peptides and proteins. Peptide-based cleavable tethers are generally limited to peptide bonds (i.e., amide bonds) formed between amino acids to give rise to peptides and proteins, and do not include the entire amide functionality. Peptide-based cleavable tethers have the general formula —NHCHRAC(O)NHCHRBC(O)— (SEQ ID NO: 13), where R and R are the R groups of two adjacent amino acids. These candidates can be evaluated using methods similar to those described above.

[0225] The ASGPR-binding moiety can be attached to any nucleotide position within a domain of a TREM (ASt domain 1, DH domain, ACH domain, VL domain, TH domain, and / or ASt domain 2). In some embodiments, the ASGPR moiety is attached to a nucleobase, terminus, or internucleotide linkage within a TREM. In some embodiments, the ASGPR moiety is attached to a nucleobase within a TREM. In some embodiments, the ASGPR-binding moiety is attached to any adenine nucleobase within a domain of a TREM (ASt domain 1, DH domain, ACH domain, VL domain, TH domain, and / or ASt domain 2). In some embodiments, the ASGPR-binding moiety is attached to any cytosine nucleobase within a domain of a TREM (ASt domain 1, DH domain, ACH domain, VL domain, TH domain, and / or ASt domain 2). In some embodiments, it is attached to any guanosine nucleobase within a domain of a TREM (ASt domain 1, DH domain, ACH domain, VL domain, TH domain, and / or ASt domain 2). In some embodiments, it is attached to any uracil nucleobase within a domain of TREM (ASt domain 1, DH domain, ACH domain, VL domain, TH domain, and / or ASt domain 2). In some embodiments, it is attached to any thymine nucleobase within a domain of TREM (ASt domain 1, DH domain, ACH domain, VL domain, TH domain, and / or ASt domain 2).

[0226] In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 1 (e.g., present in the nucleobase at TREM position 1). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 2 (e.g., present in the nucleobase at TREM position 2). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 3 (e.g., present in the nucleobase at TREM position 3). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 4 (e.g., present in the nucleobase at TREM position 4). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 5 (e.g., present in the nucleobase at TREM position 5). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 6 (e.g., present in the nucleobase at TREM position 6). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 7 (e.g., present in the nucleobase at TREM position 7). In certain embodiments, the ASGPR binding moiety is present within the TREM at TREM position 8 (e.g., is present within the nucleobase at TREM position 8). In certain embodiments, the ASGPR binding moiety is present within the TREM at TREM position 9 (e.g., is present within the nucleobase at TREM position 9).

[0227] In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 10 (e.g., present within the nucleobase at TREM position 10). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 11 (e.g., present within the nucleobase at TREM position 11). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 12 (e.g., present within the nucleobase at TREM position 12). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 13 (e.g., present within the nucleobase at TREM position 13). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 14 (e.g., present within the nucleobase at TREM position 14). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 15 (e.g., present within the nucleobase at TREM position 15). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 16 (e.g., present within the nucleobase at TREM position 16). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 17 (e.g., present within the nucleobase at TREM position 17). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 18 (e.g., present within the nucleobase at TREM position 18). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 19 (e.g., present within the nucleobase at TREM position 19). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 20 (e.g., present within the nucleobase at TREM position 20). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 21 (e.g., present within the nucleobase at TREM position 21). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 22 (e.g., present within the nucleobase at TREM position 22). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 23 (e.g., present within the nucleobase at TREM position 23).In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 24 (e.g., present within the nucleobase at TREM position 24). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 25 (e.g., present within the nucleobase at TREM position 25). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 26 (e.g., present within the nucleobase at TREM position 26).

[0228] In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 27 (e.g., present in the nucleobase at TREM position 27). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 28 (e.g., present in the nucleobase at TREM position 28). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 29 (e.g., present in the nucleobase at TREM position 29). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 30 (e.g., present in the nucleobase at TREM position 30). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 31 (e.g., present in the nucleobase at TREM position 31). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 32 (e.g., present in the nucleobase at TREM position 32). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 33 (e.g., present in the nucleobase at TREM position 33). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 34 (e.g., present in the nucleobase at TREM position 34). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 35 (e.g., present in the nucleobase at TREM position 35). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 36 (e.g., present in the nucleobase at TREM position 36). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 37 (e.g., present in the nucleobase at TREM position 37). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 38 (e.g., present in the nucleobase at TREM position 38). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 39 (e.g., present in the nucleobase at TREM position 39). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 40 (e.g., present in the nucleobase at TREM position 40).In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 41 (e.g., within the nucleobase at TREM position 41). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 42 (e.g., within the nucleobase at TREM position 42). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 43 (e.g., within the nucleobase at TREM position 43).

[0229] In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 44 (e.g., present in the nucleobase at TREM position 44). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 45 (e.g., present in the nucleobase at TREM position 45). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 46 (e.g., present in the nucleobase at TREM position 46). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 47 (e.g., present in the nucleobase at TREM position 47). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 48 (e.g., present in the nucleobase at TREM position 48). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 49 (e.g., present in the nucleobase at TREM position 49).

[0230] In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 50 (e.g., present in the nucleobase at TREM position 50). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 51 (e.g., present in the nucleobase at TREM position 51). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 52 (e.g., present in the nucleobase at TREM position 52). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 53 (e.g., present in the nucleobase at TREM position 53). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 54 (e.g., present in the nucleobase at TREM position 54). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 55 (e.g., present in the nucleobase at TREM position 55). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 56 (e.g., present in the nucleobase at TREM position 56). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 57 (e.g., present in the nucleobase at TREM position 57). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 58 (e.g., present in the nucleobase at TREM position 58). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 59 (e.g., present in the nucleobase at TREM position 59). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 60 (e.g., present in the nucleobase at TREM position 60). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 61 (e.g., present in the nucleobase at TREM position 61). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 62 (e.g., present in the nucleobase at TREM position 62). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 63 (e.g., present in the nucleobase at TREM position 63).In certain embodiments, the ASGPR binding moiety is present within the TREM at TREM position 64 (eg, is present within the nucleobase at TREM position 64).

[0231] In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 65 (e.g., present in the nucleobase at TREM position 65). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 66 (e.g., present in the nucleobase at TREM position 66). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 67 (e.g., present in the nucleobase at TREM position 67). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 68 (e.g., present in the nucleobase at TREM position 68). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 69 (e.g., present in the nucleobase at TREM position 69). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 70 (e.g., present in the nucleobase at TREM position 70). In some embodiments, the ASGPR binding moiety is present in the TREM at TREM position 71 (e.g., present in the nucleobase at TREM position 71). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 72 (e.g., present within the nucleobase at TREM position 72). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 73 (e.g., present within the nucleobase at TREM position 73). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 74 (e.g., present within the nucleobase at TREM position 74). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 75 (e.g., present within the nucleobase at TREM position 75). In some embodiments, the ASGPR binding moiety is present within the TREM at TREM position 76 (e.g., present within the nucleobase at TREM position 76).

[0232] In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 1 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 2 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 3 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 4 (U). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 5 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 6 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 7 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 8 (U). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 9 (G).

[0233] In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 10 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 11 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 12 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 13 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 14 (A). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 15 (A). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 16 (U). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 17 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 18 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 19 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 20 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 21 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 22 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 23 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 24 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 25 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 26 (A).

[0234] In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 27 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 28 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 29 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 30 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 31 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 32 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 33 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 34 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 35 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 36 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 37 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 38 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 39 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 40 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 41 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 42 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 43 (A).

[0235] In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 44 (A). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 45 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 46 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 47 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 48 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 49 (C).

[0236] In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 50 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 51 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 52 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 53 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 54 (U). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 55 (U). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 56 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 57 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 58 (A). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 59 (G). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 60 (U). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 61 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 62 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 63 (C). In some embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 64 (G).

[0237] In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 76 (A). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 75 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 74 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 73 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 72 (C). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 71 (U). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 70 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 69 (A). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 68 (G). In some embodiments, the ASGPR binding moiety is attached to a nucleobase at TREM position 67 (G). In certain embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 66 (C). In certain embodiments, the ASGPR binding moiety is attached to the nucleobase at TREM position 65 (G).

[0238] In certain embodiments, a TREM comprising an ASGPR-binding moiety comprises a deoxyribonucleic acid (DNA) sequence disclosed in Table 1, e.g., a ribonucleic acid (RNA) sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In certain embodiments, a TREM comprising an ASGPR-binding moiety comprises an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM comprising the ASGPR binding moiety comprises an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0239] In some embodiments, a TREM comprising an ASGPR-binding moiety comprises at least 5, 10, 15, 20, 25, or 30 contiguous nucleotides of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., at least 5, 10, 15, 20, 25, or 30 contiguous nucleotides of an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises at least 5, 10, 15, 20, 25, or 30 contiguous nucleotides of an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM comprising the ASGPR binding moiety comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0240] In one embodiment, the TREM comprising the ASGPR binding moiety comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM comprising an ASGPR binding moiety comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a DNA sequence provided in Table 1, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM comprising an ASGPR binding moiety comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0241] In certain embodiments, a TREM comprising an ASGPR-binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt, or 60 nt (but less than the full length) of a DNA sequence disclosed in Table 1, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In certain embodiments, a TREM comprising an ASGPR-binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt, or 60 nt (but less than the full length) of an RNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a DNA sequence provided in Table 1, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In one embodiment, the TREM comprising the ASGPR binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence having at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identity to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0242] In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a deoxyribonucleic acid (DNA) sequence disclosed in Table 4, e.g., a ribonucleic acid (RNA) sequence encoded by any one of SEQ ID NOs: 452-561 disclosed in Table 4. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 4, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 452-561 disclosed in Table 4. In one embodiment, the TREM comprising the ASGPR binding moiety comprises an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 4, e.g., any one of SEQ ID NOs: 452-561 disclosed in Table 4.

[0243] In one embodiment, the TREM comprising the ASGPR binding portion comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence provided in Table 4, e.g., any one of SEQ ID NOs: 452-561 disclosed in Table 4. In one embodiment, the TREM comprising the ASGPR binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a DNA sequence provided in Table 4, e.g., an RNA sequence encoded by any one of SEQ ID NOs: 452-561 disclosed in Table 4. In one embodiment, the TREM comprising the ASGPR binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence having at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identity to a DNA sequence provided in Table 4, e.g., any one of SEQ ID NOs: 452-561 disclosed in Table 4.

[0244] In certain embodiments, the TREM comprising an ASGPR binding moiety is any one of the compounds provided in Table 12, e.g., compound numbers 99-131. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 99. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 100. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 101. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 102. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 103. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 104. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 105. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 106. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 107. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 108. In certain embodiments, the TREM comprising an ASGPR binding moiety is compound 109. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 110. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 111. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 112. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 113. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 114. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 115. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 116. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 117. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 118. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 119. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 120. In one embodiment, a TREM comprising an ASGPR binding moiety is Compound 121. In one embodiment, the TREM comprising an ASGPR binding moiety is compound 122. In one embodiment, the TREM comprising an ASGPR binding moiety is compound 123.In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 124. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 125. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 126. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 127. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 128. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 129. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 130. In certain embodiments, a TREM comprising an ASGPR binding moiety is compound 131.

[0245] In certain embodiments, a TREM comprising an ASGPR-binding moiety comprises a compound having an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to the RNA sequence of any one of compounds 100-131 provided in Table 12. In certain embodiments, a TREM comprising an ASGPR-binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt, or 60 nt (but less than the full length) of a TREM provided in Table 12, for example, any one of compounds 100-131 disclosed in Table 12. In one embodiment, the TREM comprising the ASGPR binding portion comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of a TREM provided in Table 12, e.g., a TREM at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of compounds 100-131 disclosed in Table 12.

[0246] In certain embodiments, a TREM comprising an ASGPR binding moiety comprises any one of the sequences provided in Table 12, e.g., SEQ ID NOs: 622-654. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 622. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 623. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 624. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 625. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 626. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 627. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 628. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 629. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 630. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 631. In certain embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 632. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 633. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 634. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 635. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 636. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 637. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 638. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 639. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 640. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 641. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 642. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 643. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 644. In one embodiment, the TREM comprising an ASGPR binding portion comprises SEQ ID NO:645.In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 646. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 647. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 648. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 649. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 650. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 651. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 652. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 653. In some embodiments, a TREM comprising an ASGPR binding moiety comprises SEQ ID NO: 654.

[0247] In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a TREM provided in Table 12, e.g., the sequence of any one of SEQ ID NOs: 622-654 provided in Table 12. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt, or 60 nt (but less than the full length) of a TREM provided in Table 12, e.g., any one of SEQ ID NOs: 622-654 disclosed in Table 12. In one embodiment, the TREM comprising the ASGPR binding portion comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of a TREM provided in Table 12, for example, a TREM at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 622-654 disclosed in Table 12.

[0248] In one embodiment, the TREM comprising the ASGPR binding moiety comprises a TREM provided in Table 12, for example, a sequence that differs by 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, or 20 nt or less from any one of SEQ ID NOs: 622-652 provided in Table 12.

[0249] In some embodiments, a TREM comprising an ASGPR binding moiety is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 622. In some embodiments, a TREM comprising an ASGPR binding moiety is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 650. In one embodiment, the TREM comprising an ASGPR binding portion is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:653.

[0250] In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence that differs by at least 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, 20 nt, 25 nt, 30 nt, 40 nt, 45 nt, 50 nt, 55 nt, or more from SEQ ID NO: 622. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence that differs by no more than 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, or 20 nt from SEQ ID NO: 622. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence that differs by at least 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, 20 nt, 25 nt, 30 nt, 40 nt, 45 nt, 50 nt, 55 nt, or more from SEQ ID NO: 650. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence that differs by no more than 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, or 20 nt from SEQ ID NO: 650. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence that differs by at least 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, 20 nt, 25 nt, 30 nt, 40 nt, 45 nt, 50 nt, 55 nt, or more from SEQ ID NO: 653. In some embodiments, a TREM comprising an ASGPR-binding moiety comprises a sequence that differs by no more than 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, or 20 nt from SEQ ID NO: 653.

[0251] Chemical modification TREM In some embodiments, a TREM entity (e.g., a TREM, TREM core fragment, or TREM fragment described herein) further comprises, in addition to the ASGPR-binding moiety, a chemical modification, e.g., a modification described in any one of Tables 5-9. The chemical modification can be made according to methods known in the art. In certain embodiments, the chemical modification is a modification that cells, e.g., human cells, do not make on endogenous tRNAs.

[0252] In some embodiments, the chemical modification is a modification that a cell, e.g., a human cell, can make on an endogenous tRNA, but the modification is at a position where it does not occur on a naturally occurring tRNA. In some embodiments, the chemical modification is in a domain, linker, or arm that does not naturally have such a modification. In some embodiments, the chemical modification is at a position within a domain, linker, or arm that does not naturally have such a modification. In some embodiments, the chemical modification is on a nucleotide that does not naturally have such a modification. In some embodiments, the chemical modification is on a nucleotide at a position within a domain, linker, or arm that does not naturally have such a modification.

[0253] Any of the nucleic acids featured in this disclosure can be synthesized and / or modified by methods well established in the art, such as those described in "Current Protocols in Nucleic Acid Chemistry," Beaucage, S. Lett. et al. (Eds.), John Wiley & Sons, Inc., New York, NY, USA (incorporated herein by reference). Modifications include, for example, terminal modifications, such as 5'-terminal modifications (phosphorylation, conjugation, inverted linkage) or 3'-terminal modifications (conjugation, DNA nucleotides, inverted linkages, etc.); base modifications, such as substitution with stabilizing bases, destabilizing bases, or bases that base-pair with a wider range of partners, base removal (abasic nucleotides), or conjugated bases; sugar modifications (e.g., at the 2' or 4' position) or sugar substitutions; and / or backbone modifications, including modification or substitution of phosphodiester linkages. Specific examples of TREM compounds useful in the embodiments described herein include, but are not limited to, TREMs that contain modified backbones or do not contain natural internucleoside linkages. TREMs with modified backbones include, in particular, those that do not have a phosphorus atom in the backbone. For the purposes of this specification, and as sometimes referred to in the art, modified RNAs that do not have a phosphorus atom in the internucleoside backbone can also be considered oligonucleosides. In some embodiments, modified TREMs will have a phosphorus atom in their internucleoside backbone.

[0254] Modified TREM scaffolds include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methylphosphonates, and other alkyl phosphonates, including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates, including 3'-aminophosphoramidates and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates with normal 3'-5' linkages, 2'-5' linked analogs thereof, and those with inverted polarity, where adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed salts, and free acid forms are also included.

[0255] Representative United States patents disclosing the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S. Patent Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,195; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321 ,131 specification; 5,399,676 specification; 5,405,939 specification; 5,453,496 specification; 5,455,233 specification; 5,466,677 specification; 5,476,925 specification ; Specification No. 5,519,126; Specification No. 5,536,821; Specification No. 5,541,316; Specification No. 5,550,111; Specification No. 5,563,253; Specification No. 5,571,799; Specification No. 5,587, Specification No. 361; Specification No. 5,625,050; Specification No. 6,028,188; Specification No. 6,124,445; Specification No. 6,160,109; Specification No. 6,169,170; Specification No. 6,172,209; Specification No. 6,239,265; Specification No. 6,277,603; Specification No. 6,326,199; Specification No. 6,346,614; Specification No. 6,444,423; Specification No. 6,531,590; Specification No. 6,534,6 Nos. 39; 6,608,035; 6,683,167; 6,858,715; 6,867,294; 6,878,805; 7,015,315; 7,041,816; 7,273,933; 7,321,029; and U.S. Reissue Patent No. RE39464, the entire contents of each of which are incorporated herein by reference.

[0256] Modified TREM backbones that do not contain internal phosphorus atoms have backbones formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatom or heterocyclic internucleoside linkages. These include those with morpholino linkages (formed in part from the sugar portion of the nucleoside), siloxane backbones, sulfide, sulfoxide, and sulfone backbones, formacetyl and thioformacetyl backbones, methyleneformacetyl and thioformacetyl backbones, alkene-containing backbones, sulfamate backbones, methyleneimino and methylenehydrazino backbones, sulfonate and sulfonamide backbones, amide backbones, and others with mixed N, O, S, and CH constituent moieties. Representative United States patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Patent Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,64,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; Nos. 89,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, the entire contents of each of which are incorporated herein by reference.

[0257] In other embodiments, suitable RNA mimics are contemplated for use in TREM, in which both the sugar and internucleoside linkages, i.e., the backbone of the nucleotide units, are replaced with novel groups. The base units are maintained for hybridization with appropriate nucleic acid target compounds. One such oligomeric compound, an RNA mimic, that has been shown to have excellent hybridization properties is called peptide nucleic acid (PNA). In PNA compounds, the sugar backbone of RNA is replaced with an amide-containing backbone, particularly an aminoethylglycine backbone. The nucleobases are retained and are directly or indirectly linked to the aza nitrogen atoms of the amide portion of the backbone. Representative U.S. patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are incorporated herein by reference. Additional PNA compounds suitable for use in the TREM of the present disclosure are described, for example, in Nielsen et al., Science, 1991, 254, 1497-1500.

[0258] Some embodiments featured in this disclosure include TREMs with phosphorothioate backbones and oligonucleosides with heteroatom backbones, particularly -CH-NH-CH-, -CH-N(CH)-O-CH- (known as methylene(methylimino) or MMI backbones), -CH-O-N(CH)-CH-, -CH-N(CH)-N(CH)-CH-, and -N(CH)-CH-CH- (where the natural phosphodiester backbone is represented as -OPO-CH-) of U.S. Pat. No. 5,489,677, as described above, and the amide backbones of U.S. Pat. No. 5,602,240, as described above. In some embodiments, TREMs featured herein have the morpholino backbone structure of U.S. Pat. No. 5,034,506, as described above.

[0259] The TREMs featured herein can include one of the following at the 2' position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S-, or N-alkynyl; or O-alkyl-O-alkyl, where alkyl, alkenyl, and alkynyl are substituted or unsubstituted Ci-C 10 Alkyl or C2-C 10 It can be alkenyl and alkynyl. Exemplary suitable modifications include O[(CH) n O] m CH3, O(CH2). n OCH3, O(CH2) n NH2, O(CH2) n CH3, O(CH2) n ONH2 and 0(CH2) n ON[(CH2) n CH3)]2, where n and m are from 1 to about 10. In other embodiments, the TREM is at the 2' position and 10 It may include one of lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ON02, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, RNA cleaving group, reporter group, intercalator, group for improving the pharmacokinetic properties of TREM, or group for improving the pharmacodynamic properties of TREM, and other substituents with similar properties.

[0260] In some embodiments, the modification comprises 2'-methoxyethoxy (2'-O-CH2CHOCH3, also known as 2'-O-(2-methoxyethyl) or 2'-MOE) (Martin et al., Helv. Chim. Acta, 1995, 78:486-504), i.e., an alkoxy-alkoxy group. Another exemplary modification is 2'-dimethylaminooxyethoxy, also known as 2'-DMAOE, i.e., the O(CH2)2ON(CH3)2 group, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylaminoethoxyethyl or 2'-DMAEOE), i.e., 2'-O-CH2-O-CH2-N(CH2), as described in the Examples herein below.

[0261] Other modifications include 2'-methoxy (2'-OCH), 2'-aminopropoxy (2'-OCHCHCHNH), and 2'-fluoro (2'-F). Similar modifications can also be made at other positions within a TREM, particularly the 3' position of the sugar on the 3'-terminal nucleotide or in 2'-5'-linked TREMs and the 5' position of the 5'-terminal nucleotide. TREMs can also have sugar mimetics such as a cyclobutyl moiety in place of the pentofuranosyl sugar. Representative United States patents that teach the preparation of such modified sugar structures include, but are not limited to, U.S. Patent Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567, Nos. 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, several of which are co-owned with the present application, the entire contents of each of which are incorporated herein by reference.

[0262] TREMs may also include nucleobase (often simply referred to in the art as "base") modifications or substitutions. As used herein, "unmodified" or "natural" nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleobases include deoxythymine (dT), 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyluracil and cytosine, 6-azouracil, cytosine and thymine, 5-uracil (pseudouracil), and cytosine. Other synthetic and natural nucleobases include uracil, 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.Further nucleobases include those disclosed in U.S. Patent No. 3,687,808, those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008; those disclosed in The Concise Encyclopedia of Polymer Science and Engineering, pages 858-859, Kroschwitz, JL, ed. John Wiley & Sons, 1990, those disclosed by Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613, and those disclosed by Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, ST and Lebleu, B., Ed., CRC Press, 1993. Some of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds featured in the present invention. These include 5-substituted pyrimidines, 6-azapyrimidines, and N-2, N-6, and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil, and 5-propynylcytosine. 5-Methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2°C (Sanghvi, YS, Crooke, ST, and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are exemplary base substitutions, especially when combined with 2'-O-methoxyethyl sugar modifications.

[0263] Representative United States patents that teach the preparation of some of the above-mentioned modified nucleobases, as well as other modified nucleobases, include, but are not limited to, the above-mentioned U.S. Patent Nos. 3,687,808; 4,845,205; 5,130,30; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121; , 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200; 6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610; 7,427,672; and 7,495,088, the entire contents of each of which are incorporated herein by reference.

[0264] TREMs can also be modified to contain one or more bicyclic sugar moieties. A "bicyclic sugar" is a furanosyl ring modified by a two-atom bridge. A "bicyclic nucleoside" ("BNA") is a nucleoside having a sugar moiety containing a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4'-carbon and 2'-carbon of the sugar ring. Thus, in some embodiments, agents of the present invention may include TREM RNAs, which can also be modified to contain one or more locked nucleic acids (LNAs). Locked nucleic acids are nucleotides with modified ribose moieties that contain an additional bridge connecting the 2' and 4' carbons of the ribose moiety. In other words, LNAs are nucleotides containing a bicyclic sugar moiety containing a 4'-CH2-O-2' bridge. This structure effectively "locks" the ribose in the 3'-endo structural configuration. The addition of locked nucleic acids to oligonucleotide sequences has been shown to increase their stability in serum and reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research 33(1):439-447; Mook, OR. et al., (2007) Mol Cane Ther 6(3):833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31(12):3185-3193).

[0265] In certain embodiments, the TREM, TREM core fragment or TREM fragment described herein comprises a chemical modification provided in Table 5, or a combination thereof.

[0266] [Table 25]

[0267] [Table 26]

[0268] [Table 27]

[0269] [Table 28]

[0270] [Table 29]

[0271] [Table 30]

[0272] [Table 31]

[0273] [Table 32]

[0274] [Table 33]

[0275] [Table 34]

[0276] In some embodiments, the TREM, TREM core fragment or TREM fragment described herein comprises a modification, or a combination thereof, provided in Table 6. The modifications provided in Table 6 occur naturally in RNA and are used herein on synthetic TREM, TREM core fragment or TREM fragment at non-naturally occurring positions.

[0277] [Table 35]

[0278] [Table 36]

[0279] [Table 37]

[0280] In certain embodiments, the TREM, TREM core fragment or TREM fragment described herein comprises a chemical modification provided in Table 7, or a combination thereof.

[0281] [Table 38]

[0282] [Table 39]

[0283] [Table 40]

[0284] In certain embodiments, the TREM, TREM core fragment or TREM fragment described herein comprises a chemical modification provided in Table 8, or a combination thereof.

[0285] [Table 41]

[0286] [Table 42]

[0287] In certain embodiments, a TREM, TREM core fragment or TREM fragment described herein comprises a non-naturally occurring modification provided in Table 9, or a combination thereof.

[0288] [Table 43]

[0289] TREM, TREM core fragments and TREM fragment fusions In some embodiments, the TREM, TREM core fragment, and TREM fragment disclosed herein comprise an additional moiety, such as a fusion moiety. In some embodiments, the fusion moiety may be used to alter the folding of the TREM, TREM core fragment, and TREM fragment for purification or may be used as a targeting moiety. In some embodiments, the fusion moiety may comprise a tag, a linker, be cleavable, or contain a binding site for an enzyme. In some embodiments, the fusion moiety may be located at the N-terminus of the TREM, TREM core fragment, and TREM fragment, or at the C-terminus of the TREM, TREM core fragment, and TREM fragment. In some embodiments, the fusion moiety may be encoded by the same or different nucleic acid molecules encoding the TREM, TREM core fragment, and TREM fragment.

[0290] TREM consensus sequence In certain embodiments, the TREMs disclosed herein comprise the consensus sequences provided herein.

[0291] In certain embodiments, the TREM disclosed herein has formula I ZZZ The consensus sequence of ( ZZZ represents any of the 20 amino acids, and Formula I corresponds to all species).

[0292] In certain embodiments, the TREM disclosed herein has the formula II ZZZ The consensus sequence of ( ZZZ represents any of the 20 amino acids, and Formula II corresponds to mammals).

[0293] In certain embodiments, the TREM disclosed herein has formula III ZZZ The consensus sequence of ( ZZZrepresents any of the 20 amino acids, and Formula III corresponds to the human form).

[0294] In one embodiment, ZZZ represents any of the 20 amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.

[0295] In one embodiment, the TREM disclosed herein comprises: a) under physiological conditions, residue R0 forms a linker region, e.g., linker 1 region; b) Under physiological conditions, residues R1-R2-R3-R4-R5-R6-R7 and residues R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 forms a stem region, e.g., the AStD stem region; c) under physiological conditions, residues R8-R9 form a linker region, e.g., linker 2 region; d) Under physiological conditions, residue -R 10 -R 11 -R 12 -R 13 -R 14 R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 forms a stem-loop region, e.g., a D-arm region; e) Under physiological conditions, residue -R 29forms a linker region, e.g., linker 3 region; f) Under physiological conditions, residue -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 forms a stem-loop region, e.g., an AC arm region; g) Under physiological conditions, residues -[R 47 ] x comprises a variable region, e.g., as described herein; h) Under physiological conditions, residue -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 forms a stem-loop region, e.g., a T-arm region; or i) Under physiological conditions, residue R 72 forms a linker region, e.g., linker 4 region The characteristic includes a property selected from:

[0296] Alanine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I ALA (SEQ ID NO: 562) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10-R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Ala is R0 = absent; R 14 , R 57 = independently A or absent; R 26 = A, C, G or absent; R5, R6, R 15 , R 16 , R 21 , R 30 , R 31 , R 32 , R 34 , R 37 , R 41 , R 42 , R 43 , R 44 , R 45 , R 48 , R 49 , R 50 , R 58 , R 59 , R 63 , R 64 , R 66 , R 67 = independently N or absent; R 11 , R 35 , R 65 = independently A, C, U or absent; R1, R9, R 20 , R 38 , R 40 , R 51 , R 52 , R 56 = independently A, G or absent; R7, R 22 , R 25 , R 27 , R 29 , R 46 , R 53 , R 72 = independently A, G, U or absent; R 24 , R 69 = independently A, U or absent; R 70 , R 71 = independently C or absent; R3, R4 = independently C, G, or absent; R 12, R 33 , R 36 , R 62 , R 68 = independently C, G, U or absent; R 13 , R 17 , R 28 , R 39 , R 55 , R 60 , R 61 = independently C, U or absent; R 10 , R 19 , R 23 = independently G or absent; R2 = G, U or absent; R8, R 18 , R 54 = independently U or absent; [R 47 ] x =N or absent; Here, for example, x = 1 to 271 (e.g., x = 1 to 250, x = 1 to 225, x = 1 to 200, x = 1 to 175, x = 1 to 150, x = 1 to 125, x = 1 to 100, x = 1 to 75, x = 1 to 50, x = 1 to 40, x = 1 to 30, x = 1 to 29, x = 1 to 28, x = 1 to 27, x = 1 to 26, x = 1 to 25, x = 1 to 24, x = 1 to 2 3, x=1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~ 12, x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~ 271, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x= 1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x= 18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0297] In certain embodiments, the TREM disclosed herein has the formula II ALA (SEQ ID NO: 563), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Ala is R0, R 18 = non-existent; R 14 , R 24 , R 57 = independently A or absent; R 15 , R 26 , R64 = independently A, C, G or absent; R 16 , R 31 , R 50 , R 59 = independently N or absent; R 11 , R 32 , R 37 , R 41 , R 43 , R 45 , R 49 , R 65 , R 66 = independently A, C, U or absent; R1, R5, R9, R 25 , R 27 , R 38 , R 40 , R 46 , R 51 , R 56 = independently A, G or absent; R7, R 22 , R 29 , R 42 , R 44 , R 53 , R 63 , R 72 = independently A, G, U or absent; R6, R 35 , R 69 = independently A, U or absent; R 55 , R 60 , R 70 , R 71 = independently C or absent; R3=C, G or absent; R 12 , R 36 , R 48 = independently C, G, U or absent; R 13 , R 17 , R 28 , R 30 , R 34 , R 39 , R 58 , R 61 , R 62 , R 67 , R 68= independently C, U or absent; R4, R 10 , R 19 , R 20 , R 23 , R 52 = independently G or absent; R2, R8, R 33 = independently G, U or absent; R 21 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0298] In certain embodiments, the TREM disclosed herein has formula III ALA (SEQ ID NO: 564), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Ala is R0, R 18 = non-existent; R 14 , R 24 , R 57 , R 72 = independently A or absent; R 15 , R 26 , R 64 = independently A, C, G or absent; R 16 , R 31 , R 50 = independently N or absent; R 11 , R 32 , R 37 , R 41 , R 43 , R 45 , R 49 , R 65 , R 66 = independently A, C, U or absent; R5, R9, R 25 , R 27 , R 38 , R 40 , R 46 , R 51 , R 56 = independently A, G or absent; R7, R 22 , R 29 , R 42 , R 44 , R 53 , R 63 = independently A, G, U or absent; R6, R 35 = independently A, U or absent; R 55 , R 60 , R 61 , R 70 , R 71 = independently C or absent; R12 , R 48 , R 59 = independently C, G, U or absent; R 13 , R 17 , R 28 , R 30 , R 34 , R 39 , R 58 , R 62 , R 67 , R 68 = independently C, U or absent; R1, R2, R3, R4, R 10 , R 19 , R 20 , R 23 , R 52 = independently G or absent; R 33 , R 36 = independently G, U or absent; R8, R 21 , R 54 , R 69 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0299] Arginine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I ARG (SEQ ID NO: 565), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19-R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Arg is R 57 =A or absent; R9, R 27 = independently A, C, G or absent; R1, R2, R3, R4, R5, R6, R7, R 11 , R12 , R 16 , R 21 , R 22 , R 23 , R 25 , R 26 , R 29 , R 30 , R 31 , R 32 , R 33 , R 34 , R 37 , R 42 , R 44 , R 45 , R 46 , R 48 , R 49 , R 50 , R 51 , R 58 , R 62 , R 63 , R 64 , R 65 , R 66 , R 67 , R 68 , R 69 , R 70 , R 71 = independently N or absent; R 13 , R 17 , R 41 = independently A, C, U or absent; R 19 , R 20 , R 24 , R 40 , R 56 = independently A, G or absent; R 14 , R 15 , R 72 = independently A, G, U or absent; R 18 = A, U or absent; R 38 = C or absent; R 35 , R 43 , R 61 = independently C, G, U or absent; R 28 , R 55 , R 59 , R 60= independently C, U or absent; R0, R 10 , R 52 = independently G or absent; R8, R 39 = independently G, U or absent; R 36 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0300] In certain embodiments, the TREM disclosed herein has the formula II ARG (SEQ ID NO: 566) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Arg is R 18 = non-existent; R 24 , R 57 = independently A or absent; R 41 = A, C or absent; R3, R7, R 34 , R 50 = independently A, C, G or absent; R2, R5, R6, R 12 , R 26 , R 32 , R 37 , R 44 , R 58 , R 66 , R 67 , R 68 , R 70 = independently N or absent; R 49 , R 71 = independently A, C, U or absent; R1, R 15 , R 19 , R 25 , R 27 , R 40 , R 45 , R 46 , R 56 , R 72 = independently A, G or absent; R 14 , R 29 , R 63 = independently A, G, U or absent; R 16 , R 21 = independently A, U or absent; R 38 , R 61 = independently C or absent; R 33 , R 48 = independently C, G, or absent; R4, R9, R 11, R 43 , R 62 , R 64 , R 69 = independently C, G, U or absent; R 13 , R 22 , R 28 , R 30 , R 31 , R 35 , R 55 , R 60 , R 65 = independently C, U or absent; R0, R 10 , R 20 , R 23 , R 51 , R 52 = independently G or absent; R8, R 39 , R 42 = independently G, U or absent; R 17 , R 36 , R 53 , R 54 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0301] In certain embodiments, the TREM disclosed herein has formula III ARG (SEQ ID NO: 567) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Arg is R 18 = non-existent; R 15 , R 21 , R 24 , R 41 , R 57 = independently A or absent; R 34 , R44 = independently A, C or absent; R3, R5, R 58 = independently A, C, G or absent; R2, R6, R 66 , R 70 = independently N or absent; R 37 , R 49 = independently A, C, U or absent; R1, R 25 , R 29 , R 40 , R 45 , R 46 , R 50 = independently A, G or absent; R 14 , R 63 , R 68 = independently A, G, U or absent; R 16 = A, U or absent; R 38 , R 61 = independently C or absent; R7, R 11 , R 12 , R 26 , R 48 = independently C, G, or absent; R 64 , R 67 , R 69 = independently C, G, U or absent; R4, R 13 , R 22 , R 28 , R 30 , R 31 , R 35 , R 43 , R 55 , R 60 , R 62 , R 65 , R 71 = independently C, U or absent; R0, R 10 , R 19 , R 20 , R 23 , R 27 , R 33 , R 51 , R52 , R 56 , R 72 = independently G or absent; R8, R9, R 32 , R 39 , R 42 = independently G, U or absent; R 17 , R 36 , R 53 , R 54 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0302] Asparagine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I ASN The sequence (SEQ ID NO: 568): R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Asn is R0, R 18 = non-existent; R 41 =A or absent; R 14 , R 48 , R 56 = independently A, C, G or absent; R2, R4, R5, R6, R 12 , R 17 , R 26 , R 29 , R 30 , R 31 , R 44 , R 45 , R 46 , R 49 , R 50 , R 58 , R 62 , R 63 , R 65 , R 66 , R 67 , R 68 , R 70 , R 71 = independently N or absent; R 11 , R 13 , R 22 , R 42 , R 55 , R 59 = independently A, C, U or absent; R9, R 15 , R 24 , R 27 , R 34 , R 37 , R 51 , R 72 = independently A, G or absent; R1, R7, R 25 , R 69 = independently A, G, U or absent; R40 , R 57 = independently A, U or absent; R 60 = C or absent; R 33 = C, G or absent; R 21 , R 32 , R 43 , R 64 = independently C, G, U or absent; R3, R 16 , R 28 , R 35 , R 36 , R 61 = independently C, U or absent; R 10 , R 19 , R 20 , R 52 = independently G or absent; R 54 = G, U or absent; R8, R 23 , R 38 , R 39 , R 53 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0303] In certain embodiments, the TREM disclosed herein has the formula II ASN (SEQ ID NO: 569), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Asn is R0, R 18 = non-existence, R 24 , R 41 , R 46 , R 62 = independently A or absent; R 59 = A, C or absent; R 14 , R 56 , R 66 = independently A, C, G or absent; R 17 , R 29 = independently N or absent; R 11 , R 26 , R 42 , R 55 = independently A, C, U or absent; R1, R9, R 12 , R 15 , R 25 , R 34 , R 37 , R 48 , R 51 , R 67 , R 68 , R 69 , R 70 , R 72 = independently A, G or absent; R 44 , R 45 , R 58 = independently A, G, U or absent; R 40 , R 57 = independently A, U or absent; R5, R 28 , R 60 = independently C or absent; R 33 , R 65 = independently C, G, or absent; R 21 , R 43 , R 71 = independently C, G, U or absent; R3, R6, R 13 , R 22 , R 32 , R 35 , R 36 , R 61 , R 63 , R 64 = independently C, U or absent; R7, R 10 , R 19 , R 20 , R 27 , R49 , R 52 = independently G or absent; R 54 = G, U or absent; R2, R4, R8, R 16 , R 23 , R 30 , R 31 , R 38 , R 39 , R 50 , R 53 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0304] In certain embodiments, the TREM disclosed herein has formula III ASN (SEQ ID NO: 570), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Asn is R0, R 18 = non-existence, R 24 , R 40 , R 41 , R 46 , R 62 = independently A or absent; R 59 = A, C or absent; R 14 , R 56 , R 66 = independently A, C, G or absent; R 11 , R 26 , R 42 , R 55 = independently A, C, U or absent; R1, R9, R 12 , R 15 , R 34 , R 37 , R 48 , R 51 , R 67 , R 68 , R 69 , R 70 = independently A, G or absent; R 44 , R 45 , R 58 = independently A, G, U or absent; R 57 = A, U or absent; R5, R 28 , R 60 = independently C or absent; R 33 , R 65 = independently C, G, or absent; R 17 , R 21 , R 29 = independently C, G, U or absent; R3, R6, R13 , R 22 , R 32 , R 35 , R 36 , R 43 , R 61 , R 63 , R 64 , R 71 = independently C, U or absent; R7, R 10 , R 19 , R 20 , R 25 , R 27 , R 49 , R 52 , R 72 = independently G or absent; R 54 = G, U or absent; R2, R4, R8, R 16 , R 23 , R 30 , R 31 , R 38 , R 39 , R 50 , R 53 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0305] Aspartic acid TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I ASP (SEQ ID NO: 571), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19-R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Asp is R0 = Absent; R 24 , R 71 = independently A, C or absent; R 33 , R 46= independently A, C, G or absent; R2, R3, R4, R5, R6, R 12 , R 16 , R 22 , R 26 , R 29 , R 31 , R 32 , R 44 , R 48 , R 49 , R 58 , R 63 , R 64 , R 66 , R 67 , R 68 , R 69 = independently N or absent; R 13 , R 21 , R 34 , R 41 , R 57 , R 65 = independently A, C, U or absent; R9, R 10 , R 14 , R 15 , R 20 , R 27 , R 37 , R 40 , R 51 , R 56 , R 72 = independently A, G or absent; R7, R 25 , R 42 = independently A, G, U or absent; R 39 = C or absent; R 50 , R 62 = independently C, G, or absent; R 30 , R 43 , R 45 , R 55 , R 70 = independently C, G, U or absent; R8, R 11 , R 17 , R 18 , R 28 , R 35 , R 53 , R59 , R 60 , R 61 = independently C, U or absent; R 19 , R 52 = independently G or absent; R1 = G, U or absent; R 23 , R 36 , R 38 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0306] In certain embodiments, the TREM disclosed herein has the formula II ASP (SEQ ID NO: 572), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Asp is R0, R 17 , R 18 , R 23 = independently non-existent; R9, R 40 = independently A or absent; R 24 , R 71 = independently A, C or absent; R 67 , R 68 = independently A, C, G or absent; R2, R6, R 66 = independently N or absent; R 57 , R 63 = independently A, C, U or absent; R 10 , R 14 , R 27 , R 33 , R 37 , R 44 , R 46 , R 51 , R 56 , R 64 , R 72 = independently A, G or absent; R7, R 12 , R 26 , R 65 = independently A, U or absent; R 39 , R 61 , R 62 = independently C or absent; R3, R 31 , R 45 , R 70 = independently C, G, or absent; R4, R5, R 29 , R 43 , R 55 = independently C, G, U or absent; R8, R11 , R 13 , R 30 , R 32 , R 34 , R 35 , R 41 , R 48 , R 53 , R 59 , R 60 = independently C, U or absent; R 15 , R 19 , R 20 , R 25 , R 42 , R 50 , R 52 = independently G or absent; R1, R 22 , R 49 , R 58 , R 69 = independently G, U or absent; R 16 , R 21 , R 28 , R 36 , R 38 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0307] In certain embodiments, the TREM disclosed herein has formula III ASP (SEQ ID NO: 573), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Asp is R0, R 17 , R 18 , R 23 = non-existence, R9, R 12 , R 40 , R 65 , R 71 = independently A or absent; R2, R 24 , R 57 = independently A, C or absent; R6, R 14 , R 27 , R 46 , R 51 , R 56 , R 64 , R 67 , R 68 = independently A, G or absent; R3, R 31 , R 35 , R 39 , R 61 , R 62 = independently C or absent; R 66 = C, G or absent; R5, R8, R 29 , R 30 , R 32 , R 34 , R 41 , R 43 , R 48 , R 55 , R 59 , R 60 , R 63 = independently C, U or absent; R 10 , R 15 , R 19 , R 20 , R 25 , R 33 , R 37 , R 42 , R 44 , R 45 , R 49 , R 50 , R 52 , R 69 , R 70 , R 72 = independently G or absent; R 22 , R 58 = independently G, U or absent; R1, R4, R7, R 11 , R 13 , R 16 , R 21 , R 26 , R 28 , R 36 , R38 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0308] Cysteine ​​TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I CYS (SEQ ID NO: 574), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11-R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Cys is R0 = Absent; R 14 , R 39 , R 57 = independently A or absent; R 41 = A, C or absent; R 10 , R 15 , R 27 , R 33 , R 62 = independently A, C, G or absent; R3, R4, R5, R6, R 12 , R 13 , R 16 , R 24 , R 26 , R 29 , R 30 , R 31 , R 32 , R 34 , R 42 , R 44 , R 45 , R 46 , R 48 , R 49 , R 58 , R 63 , R 64 , R 66 , R 67 , R 68 , R 69 , R 70 = independently N or absent; R 65 = A, C, U or absent; R9, R 25 , R 37 , R 40 , R 52 , R 56 = independently A, G or absent; R7, R 20 , R 51 = independently A, G, U or absent; R 18 , R 38 , R 55 = independently C or absent; R2 = C, G or absent; R 21 , R 28 , R 43 , R50 = independently C, G, U or absent; R 11 , R 22 , R 23 , R 35 , R 36 , R 59 , R 60 , R 61 , R 71 , R 72 = independently C, U or absent; R1, R 19 = independently G or absent; R 17 = G, U or absent; R8, R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0309] In certain embodiments, the TREM disclosed herein has the formula II CYS (SEQ ID NO: 575), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Cys is R0, R 18 , R 23 = non-existent; R 14 , R 24 , R 26 , R 29 , R 39 , R 41 , R 45 , R57 = independently A or absent; R 44 = A, C or absent; R 27 , R 62 = independently A, C, G or absent; R 16 = A, C, G, U or absent; R 30 , R 70 = independently A, C, U or absent; R5, R7, R9, R 25 , R 34 , R 37 , R 40 , R 46 , R 52 , R 56 , R 58 , R 66 = independently A, G or absent; R 20 , R 51 = independently A, G, U or absent; R 35 , R 38 , R 43 , R 55 , R 69 = independently C or absent; R2, R4, R 15 = independently C, G, or absent; R 13 = C, G, U or absent; R6, R 11 , R 28 , R 36 , R 48 , R 49 , R 50 , R 60 , R 61 , R 67 , R 68 , R 71 , R 72 = independently C, U or absent; R1, R3, R 10 , R 19 , R 33 , R 63 = independently G or absent; R8, R 17 , R21 , R 64 = independently G, U or absent; R 12 , R 22 , R 31 , R 32 , R 42 , R 53 , R 54 , R 65 = independently U or absent; R 59 =U, or non-existent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0310] In certain embodiments, the TREM disclosed herein has formula III CYS (SEQ ID NO: 576), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Cys is R0, R 18 , R 23 = non-existence, R 14 , R 24 , R 26 , R 29 , R 34 , R 39 , R 41 , R 45 , R 57 , R 58 = independently A or absent; R 44 , R 70 = independently A, C or absent; R 62 = A, C, G or absent; R 16 =N or absent; R5, R7, R9, R 20 , R 40 , R 46 , R 51 , R 52 , R 56 , R 66 = independently A, G or absent; R 28 , R 35 , R 38 , R 43 , R 55 , R 67 , R 69 = independently C or absent; R4, R 15 = independently C, G, or absent; R6, R 11 , R 13 , R 30 , R 48 , R 49 , R 50 , R 60 , R 61 , R 68 , R 71 , R72 = independently C, U or absent; R1, R2, R3, R 10 , R 19 , R 25 , R 27 , R 33 , R 37 , R 63 = independently G or absent; R8, R 21 , R 64 = independently G, U or absent; R 12 , R 17 , R 22 , R 31 , R 32 , R 36 , R 42 , R 53 , R 54 , R 59 , R 65 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0311] Glutamine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I GLN (SEQ ID NO: 577), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19-R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Gln is R0, R 18 = non-existent; R 14 , R 24 , R 57 = independently A or absent; R9, R 26 , R27 , R 33 , R 56 = independently A, C, G or absent; R2, R4, R5, R6, R 12 , R 13 , R 16 , R 21 , R 22 , R 25 , R 29 , R 30 , R 31 , R 32 , R 34 , R 41 , R 42 , R 44 , R 45 , R 46 , R 48 , R 49 , R 50 , R 58 , R 62 , R 63 , R 66 , R 67 , R 68 , R 69 , R 70 = independently N or absent; R 17 , R 23 , R 43 , R 65 , R 71 = independently A, C, U or absent; R 15 , R 40 , R 51 , R 52 = independently A, G or absent; R1, R7, R 72 = independently A, G, U or absent; R3, R 11 , R 37 , R 60 , R 64 = independently C, G, U or absent; R 28 , R 35 , R 55 , R 59 , R 61 = independently C, U or absent; R 10 , R 19 , R20 = independently G or absent; R 39 = G, U or absent; R8, R 36 , R 38 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0312] In certain embodiments, the TREM disclosed herein has the formula II GLN (SEQ ID NO: 578) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Gln is R0, R 18 , R 23 = non-existence, R 14 , R 24 , R 57 = independently A or absent; R 17 , R 71 = independently A, C or absent; R 25 , R 26 , R 33 , R 44 , R 46 , R 56 , R 69 = independently A, C, G or absent; R4, R5, R 12 , R 22 , R 29 , R 30 , R 48 , R 49 , R 63 , R 67 , R 68 = independently N or absent; R 31 , R 43 , R 62 , R 65 , R 70 = independently A, C, U or absent; R 15 , R 27 , R 34 , R 40 , R 41 , R 51 , R 52 = independently A, G or absent; R2, R7, R 21 , R 45 , R 50 , R 58 , R 66 , R 72 = independently A, G, U or absent; R3, R 13 , R 32 , R 37 , R 42 , R 60 , R64 = independently C, G, U or absent; R6, R 11 , R 28 , R 35 , R 55 , R 59 , R 61 = independently C, U or absent; R9, R 10 , R 19 , R 20 = independently G or absent; R1, R 16 , R 39 = independently G, U or absent; R8, R 36 , R 38 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0313] In certain embodiments, the TREM disclosed herein has formula III GLN (SEQ ID NO: 579) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Gln is R0, R 18 , R 23 = non-existence, R 14 , R 24 , R 41 , R 57 = independently A or absent; R 17 , R71 = independently A, C or absent; R5, R 25 , R 26 , R 46 , R 56 , R 69 = independently A, C, G or absent; R4, R 22 , R 29 , R 30 , R 48 , R 49 , R 63 , R 68 = independently N or absent; R 43 , R 62 , R 65 , R 70 = independently A, C, U or absent; R 15 , R 27 , R 33 , R 34 , R 40 , R 51 , R 52 = independently A, G or absent; R2, R7, R 12 , R 45 , R 50 , R 58 , R 66 = independently A, G, U or absent; R 31 = A, U or absent; R 32 , R 44 , R 60 = independently C, G, or absent; R3, R 13 , R 37 , R 42 , R 64 , R 67 = independently C, G, U or absent; R6, R 11 , R 28 , R 35 , R 55 , R 59 , R 61 = independently C, U or absent; R9, R 10 , R 19 , R20 = independently G or absent; R1, R 21 , R 39 , R 72 = independently G, U or absent; R8, R 16 , R 36 , R 38 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0314] Glutamate TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I GLU (SEQ ID NO: 580) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Glu is R0 = absent; R 34 , R 43 , R 68 , R 69 = independently A, C, G or absent; R1, R2, R5, R6, R9, R 12 , R 16 , R 20 , R 21 , R 26 , R 27 , R 29 , R 30 , R 31 , R 32 , R 33 , R 41 , R 44 , R 45 , R 46 , R 48 , R 50 , R 51 , R 58 , R 63 , R 64 , R 65 , R 66 , R 70 , R 71 = independently N or absent; R 13 , R 17 , R 23 , R 61 = independently A, C, U or absent; R 10 , R 14 , R 24 , R 40 , R 52 , R 56 = independently A, G or absent; R7, R 15 , R 25 , R 67 , R 72 = independently A, G, U or absent; R 11 , R57 = independently A, U or absent; R 39 = C, G or absent; R3, R4, R 22 , R 42 , R 49 , R 55 , R 62 = independently C, G, U or absent; R 18 , R 28 , R 35 , R 37 , R 53 , R 59 , R 60 = independently C, U or absent; R 19 = G or absent; R8, R 36 , R 38 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0315] In certain embodiments, the TREM disclosed herein has the formula II GLU (SEQ ID NO: 581) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Glu is R0, R 18 , R 23 = non-existence, R 17 , R 40 = independently A or absent; R 26 , R 27 , R 34 , R43 , R 68 , R 69 , R 71 = independently A, C, G or absent; R1, R2, R5, R 12 , R 21 , R 31 , R 33 , R 41 , R 45 , R 48 , R 51 , R 58 , R 66 , R 70 = independently N or absent; R 44 , R 61 = independently A, C, U or absent; R9, R 14 , R 24 , R 25 , R 52 , R 56 , R 63 = independently A, G or absent; R7, R 15 , R 46 , R 50 , R 67 , R 72 = independently A, G, U or absent; R 29 , R 57 = independently A, U or absent; R 60 = C or absent; R 39 = C, G or absent; R3, R6, R 20 , R 30 , R 32 , R 42 , R 55 , R 62 , R 65 = independently C, G, U or absent; R4, R8, R 16 , R 28 , R 35 , R 37 , R 49 , R 53 , R 59 = independently C, U or absent; R10 , R 19 = independently G or absent; R 22 , R 64 = independently G, U or absent; R 11 , R 13 , R 36 , R 38 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0316] In certain embodiments, the TREM disclosed herein has formula III GLU (SEQ ID NO: 582) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Glu is R0, R 17 , R 18 , R 23 = non-existence, R 14 , R 27 , R 40 , R 71 = independently A or absent; R 44 = A, C or absent; R 43 = A, C, G or absent; R1, R 31 , R 33 , R 45 , R 51 , R 66 = independently N or absent; R 21 , R 41 = independently A, C, U or absent; R7, R 24 , R 25 , R 50 , R 52 , R 56 , R 63 , R 68 , R 70 = independently A, G or absent; R5, R 46 = independently A, G, U or absent; R 29 , R 57 , R 67 , R 72 = independently A, U or absent; R2, R 39 , R 60 = independently C or absent; R3, R 12 , R 20 , R 26 , R 34 , R 69 = independently C, G, or absent; R6, R 30 , R42 , R 48 , R 65 = independently C, G, U or absent; R4, R 16 , R 28 , R 35 , R 37 , R 49 , R 53 , R 55 , R 58 , R 61 , R 62 = independently C, U or absent; R9, R 10 , R 19 , R 64 = independently G or absent; R 15 , R 22 , R 32 = independently G, U or absent; R8, R 11 , R 13 , R 36 , R 38 , R 54 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0317] Glycine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I GLY (SEQ ID NO: 583), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R is 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a polynucleotide residue and the consensus for Gly is R0 = absent; R 24 =A or absent; R3, R9, R 40 , R 50 , R 51 = independently A, C, G or absent; R4, R5, R6, R7, R 12 , R 16 , R 21 , R 22 , R 26 , R 29 , R 30 , R 31 , R 32 , R 34 , R 41 , R 42 , R 43 , R 44 , R 45 , R 46 , R 48 , R 49 , R 58 , R 63 , R 64 , R 65 , R 66 , R 67 , R 68 = independently N or absent; R 59 = A, C, U or absent; R1, R 10 , R 14 , R 15 , R 27 , R 56 = independently A, G or absent; R 20 , R 25 = independently A, G, U or absent; R 57 , R 72 = independently A, U or absent; R 38 , R 39 , R 60 = independently C or absent; R 52 = C, G or absent; R2, R 19 , R 37 , R 54 , R 55 , R 61 , R 62 , R 69 , R 70 = independently C, G, U or absent; R 11 , R 13 , R17 , R 28 , R 35 , R 36 , R 71 = independently C, U or absent; R8, R 18 , R 23 , R 53 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0318] In certain embodiments, the TREM disclosed herein has the formula II GLY(SEQ ID NO: 584), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71-R 72 where R is a ribonucleotide residue and the consensus for Gly is R0, R 18 , R 23 = non-existence, R 24 , R 27 , R 40 , R 72 = independently A or absent; R 26 = A, C or absent; R3, R7, R 68 = independently A, C, G or absent; R5, R 30 , R 41 , R 42 , R 44 , R 49 , R 67 = independently A, C, G, U or absent; R 31 , R 32 , R 34 = independently A, C, U or absent; R9, R 10 , R 14 , R 15 , R 33 , R 50 , R 56 = independently A, G or absent; R 12 , R 16 , R 22 , R 25 , R 29 , R 46 = independently A, G, U or absent; R 57 = A, U or absent; R 17 , R 38 , R 39 , R 60 , R 61 , R 71 = independently C or absent; R6, R 52 , R 64 , R 66 = independently C, G, or absent; R2, R4, R37 , R 48 , R 55 , R 65 = independently C, G, U or absent; R 13 , R 35 , R 43 , R 62 , R 69 = independently C, U or absent; R1, R 19 , R 20 , R 51 , R 70 = independently G or absent; R 21 , R 45 , R 63 = independently G, U or absent; R8, R 11 , R 28 , R 36 , R 53 , R 54 , R 58 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0319] In certain embodiments, the TREM disclosed herein has formula III GLY (SEQ ID NO: 585) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Gly is R0, R 18 , R 23 = non-existence, R 24 , R 27 , R 40 , R 72 = independently A or absent; R 26= A, C or absent; R3, R7, R 49 , R 68 = independently A, C, G or absent; R5, R 30 , R 41 , R 44 , R 67 = independently N or absent; R 31 , R 32 , R 34 = independently A, C, U or absent; R9, R 10 , R 14 , R 15 , R 33 , R 50 , R 56 = independently A, G or absent; R 12 , R 25 , R 29 , R 42 , R 46 = independently A, G, U or absent; R 16 , R 57 = independently A, U or absent; R 17 , R 38 , R 39 , R 60 , R 61 , R 71 = independently C or absent; R6, R 52 , R 64 , R 66 = independently C, G, or absent; R 37 , R 48 , R 65 = independently C, G, U or absent; R2, R4, R 13 , R 35 , R 43 , R 55 , R 62 , R 69 = independently C, U or absent; R1, R 19 , R 20 , R 51 , R 70= independently G or absent; R 21 , R 22 , R 45 , R 63 = independently G, U or absent; R8, R 11 , R 28 , R 36 , R 53 , R 54 , R 58 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0320] Histidine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I HIS (SEQ ID NO: 586) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for His is R 23 = non-existent; R 14 , R 24 , R 57 = independently A or absent; R 72 = A, C or absent; R9, R 27 , R 43 , R 48 , R 69 = independently A, C, G or absent; R3, R4, R5, R6, R 12 , R 25 , R 26 , R 29 , R 30 , R 31 , R 34 , R 42 , R 45 , R 46 , R 49 , R 50 , R 58 , R 62 , R 63 , R 66 , R 67 , R 68 = independently N or absent; R 13 , R 21 , R 41 , R 44 , R 65 = independently A, C, U or absent; R 40 , R 51 , R 56 , R 70 = independently A, G or absent; R7, R 32 = independently A, G, U or absent; R 55 , R 60= independently C or absent; R 11 , R 16 , R 33 , R 64 = independently C, G, U or absent; R2, R 17 , R 22 , R 28 , R 35 , R 53 , R 59 , R 61 , R 71 = independently C, U or absent; R1, R 10 , R 15 , R 19 , R 20 , R 37 , R 39 , R 52 = independently G or absent; R0=G, U or absent; R8, R 18 , R 36 , R 38 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0321] In certain embodiments, the TREM disclosed herein has the formula II HIS The sequence (SEQ ID NO: 587): R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for His is R0, R 17 , R 18 , R 23 = non-existent; R7, R 12 , R 14 , R 24 , R 27 , R 45 , R 57 , R58 , R 63 , R 67 , R 72 = independently A or absent; R3=A, C, U or absent; R4, R 43 , R 56 , R 70 = independently A, G or absent; R 49 = A, U or absent; R2, R 28 , R 30 , R 41 , R 42 , R 44 , R 48 , R 55 , R 60 , R 66 , R 71 = independently C or absent; R 25 = C, G or absent; R9 = C, G, U or absent; R8, R 13 , R 26 , R 33 , R 35 , R 50 , R 53 , R 61 , R 68 = independently C, U or absent; R1, R6, R 10 , R 15 , R 19 , R 20 , R 32 , R 34 , R 37 , R 39 , R 40 , R 46 , R 51 , R 52 , R 62 , R 64 , R 69 = independently G or absent; R 16 = G, U or absent; R5, R 11 , R 21 , R 22 , R 29 , R31 , R 36 , R 38 , R 54 , R 59 , R 65 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0322] In certain embodiments, the TREM disclosed herein has formula III HIS The sequence (SEQ ID NO: 588): R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10-R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for His is R0, R 17 , R 18 , R 23 = non-existent; R7, R 12 , R 14 , R 24 , R 27 , R 45 , R 57 , R 58 , R 63 , R 67 , R 72 = independently A or absent; R3=A, C or absent; R4, R 43 , R 56 , R 70 = independently A, G or absent; R 49 = A, U or absent; R2, R 28 , R 30 , R 41 , R 42 , R 44 , R 48 , R 55 , R 60 , R 66 , R 71 = independently C or absent; R8, R9, R 26 , R 33 , R 35 , R 50 , R 61 , R 68 = independently C, U or absent; R1, R6, R 10 , R 15 , R 19 , R 20 , R 25 , R 32 , R 34 , R 37 , R 39 , R 40 , R 46 , R 51 , R 52 , R 62 , R 64 , R 69 = independently G or absent; R5, R 11 , R13 , R 16 , R 21 , R 22 , R 29 , R 31 , R 36 , R 38 , R 53 , R 54 , R 59 , R 65 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0323] Isoleucine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I ILE The sequence (SEQ ID NO: 589): R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for He is R 23 = non-existent; R 38 , R 41 , R 57 , R 72 = independently A or absent; R1, R 26 = independently A, C, G or absent; R0, R3, R4, R6, R 16 , R 31 , R 32 , R 34 , R 37 , R 42 , R 43 , R 44 , R 45 , R 46 , R 48 , R 49 , R 50 , R 58 , R 59 , R 62 , R 63 , R 64 , R 66 , R 67 , R 68 , R 69 = independently N or absent; R 22 , R 61 , R 65 = independently A, C, U or absent; R9, R 14 , R 15 , R 24 , R 27 , R 40 = independently A, G or absent; R7, R 25 , R 29 , R 51 , R 56 = independently A, G, U or absent; R 18 , R 54 = independently A, U or absent; R 60 = C or absent; R2, R 52 , R 70 = independently C, G, or absent; R5, R 12 , R 21 , R 30 , R 33 , R 71 = independently C, G, U or absent; R 11 , R 13 , R 17 , R 28 , R 35 , R 53 , R 55 = independently C, U or absent; R 10 , R 19 , R 20 = independently G or absent; R8, R 36 , R 39 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0324] In certain embodiments, the TREM disclosed herein has the formula II ILE (SEQ ID NO: 590) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for He is R0, R 18 , R 23 = non-existence, R 24 , R 38 , R 40 , R 41 , R 57 , R 72 = independently A or absent; R 26 , R 65 = independently A, C or absent; R 58 , R 59 , R 67 = independently N or absent; R 22 = A, C, U or absent; R6, R9, R 14 , R 15 , R 29 , R 34 , R 43 , R 46 , R 48 , R 50 , R 51 , R 63 , R 69 = independently A, G or absent; R 37 , R 56 = independently A, G, U or absent; R 54 = A, U or absent; R 28 , R 35 , R 60 , R 62 , R 71 = independently C or absent; R2, R 52 , R 70 = independently C, G, or absent; R5 = C, G, U or absent; R3, R4, R 11 , R 13 , R 17 , R 21 , R 30 , R 42 , R 44 , R 45 , R 49 , R 53 , R 55 , R 61 , R 64 , R 66 = independently C, U or absent; R1, R 10 , R 19 , R 20 , R 25 , R 27, R 31 , R 68 = independently G or absent; R7, R 12 , R 32 = independently G, U or absent; R8, R 16 , R 33 , R 36 , R 39 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0325] In certain embodiments, the TREM disclosed herein has formula III ILE (SEQ ID NO: 591) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for He is R0, R 18 , R 23 = non-existence, R 14 , R 24 , R 38 , R 40 , R 41 , R 57 , R 72 = independently A or absent; R 26 , R 65 = independently A, C or absent; R 22 , R 59 = independently A, C, U or absent; R6, R9, R 15 , R 34 , R 43 , R 46 , R 51 , R 56 , R 63 , R 69 = independently A, G or absent; R 37 = A, G, U or absent; R 13 , R 28 , R 35 , R 44 , R 55 , R 60 , R 62 , R 71 = independently C or absent; R2, R5, R 70 = independently C, G, or absent; R 58 , R 67 = independently C, G, U or absent; R3, R4, R 11 , R 17 , R 21 , R 30 , R 42 , R 45 , R 49 , R 53, R 61 , R 64 , R 66 = independently C, U or absent; R1, R 10 , R 19 , R 20 , R 25 , R 27 , R 29 , R 31 , R 32 , R 48 , R 50 , R 52 , R 68 = independently G or absent; R7, R 12 = independently G, U or absent; R8, R 16 , R 33 , R 36 , R 39 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0326] Methionine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I MET (SEQ ID NO: 592) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Met is R0, R 23 = non-existent; R 14 , R 38 , R 40 , R 57 = independently A or absent; R60 = A, C or absent; R 33 , R 48 , R 70 = independently A, C, G or absent; R1, R3, R4, R5, R6, R 11 , R 12 , R 16 , R 17 , R 21 , R 22 , R 26 , R 27 , R 29 , R 30 , R 31 , R 32 , R 42 , R 44 , R 45 , R 46 , R 49 , R 50 , R 58 , R 62 , R 63 , R 66 , R 67 , R 68 , R 69 , R 71 = independently N or absent; R 18 , R 35 , R 41 , R 59 , R 65 = independently A, C, U or absent; R9, R 15 , R 51 = independently A, G or absent; R7, R 24 , R 25 , R 34 , R 53 , R 56 = independently A, G, U or absent; R 72 = A, U or absent; R 37 = C or absent; R 10 , R 55 = independently C, G, or absent; R2, R 13 , R 28 , R43 , R 64 = independently C, G, U or absent; R 36 , R 61 = independently C, U or absent; R 19 , R 20 , R 52 = independently G or absent; R8, R 39 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0327] In certain embodiments, the TREM disclosed herein has the formula II MET (SEQ ID NO: 593), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Met is R0, R 18 , R 22 , R 23 = non-existence, R 14 , R 24 , R 38 , R 40 , R 41 , R 57 , R 72 = independently A or absent; R 59 , R 60 , R 62 , R 65 = independently A, C or absent; R6, R 45 , R 67 = independently A, C, G or absent; R4=N or absent; R 21 , R 42 = independently A, C, U or absent; R1, R9, R 27 , R 29 , R 32 , R 46 , R 51 = independently A, G or absent; R 17 , R 49 , R 53 , R 56 , R 58 = independently A, G, U or absent; R 63 = A, U or absent; R3, R 13 , R 37 = independently C or absent; R 48 , R 55 , R 64 , R 70 = independently C, G, or absent; R2, R5, R66 , R 68 = independently C, G, U or absent; R 11 , R 16 , R 26 , R 28 , R 30 , R 31 , R 35 , R 36 , R 43 , R 44 , R 61 , R 71 = independently C, U or absent; R 10 , R 12 , R 15 , R 19 , R 20 , R 25 , R 33 , R 52 , R 69 = independently G or absent; R7, R 34 , R 50 = independently G, U or absent; R8, R 39 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0328] In certain embodiments, the TREM disclosed herein has formula III MET (SEQ ID NO: 594) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Met is R0, R 18 , R 22 , R 23 = non-existence, R 14 , R 24 , R 38 , R 40 , R 41 , R 57 , R72 = independently A or absent; R 59 , R 62 , R 65 = independently A, C or absent; R6, R 67 = independently A, C, G or absent; R4, R 21 = independently A, C, U or absent; R1, R9, R 27 , R 29 , R 32 , R 45 , R 46 , R 51 = independently A, G or absent; R 17 , R 56 , R 58 = independently A, G, U or absent; R 49 , R 53 , R 63 = independently A, U or absent; R3, R 13 , R 26 , R 37 , R 43 , R 60 = independently C or absent; R2, R 48 , R 55 , R 64 , R 70 = independently C, G, or absent; R5, R 66 = independently C, G, U or absent; R 11 , R 16 , R 28 , R 30 , R 31 , R 35 , R 36 , R 42 , R 44 , R 61 , R 71 = independently C, U or absent; R 10 , R 12 , R 15 , R 19 , R 20, R 25 , R 33 , R 52 , R 69 = independently G or absent; R7, R 34 , R 50 , R 68 = independently G, U or absent; R8, R 39 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0329] Leucine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I LEU (SEQ ID NO: 595) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Leu is R0 = absent; R 38 , R 57 = independently A or absent; R 60 = A, C or absent; R1, R 13 , R 27 , R 48 , R 51 , R 56 = independently A, C, G or absent; R2, R3, R4, R5, R6, R7, R9, R 10 , R 11 , R 12 , R 16 , R 23 , R 26 , R 28 , R 29 , R 30 , R 31 , R 32 , R 33 , R 34 , R 37 , R 41 , R 42 , R 43 , R 44 , R 45 , R 46 , R 49 , R 50 , R 58 , R 62 , R 63 , R 65 , R 66 , R 67 , R 68 , R 69 , R 70 = independently N or absent; R 17 , R 18 , R 21 , R 22 , R 25 , R 35 , R 55= independently A, C, U or absent; R 14 , R 15 , R 39 , R 72 = independently A, G or absent; R 24 , R 40 = independently A, G, U or absent; R 52 , R 61 , R 64 , R 71 = independently C, G, U or absent; R 36 , R 53 , R 59 = independently C, U or absent; R 19 = G or absent; R 20 = G, U or absent; R8, R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0330] In certain embodiments, the TREM disclosed herein has the formula II LEU Sequence (SEQ ID NO: 596) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Leu is R0 = Absent; R 38 , R 57 , R 72 = independently A or absent; R 60 = A, C or absent; R4, R5, R 48 , R50 , R 56 , R 69 = independently A, C, G or absent; R6, R 33 , R 41 , R 43 , R 46 , R 49 , R 58 , R 63 , R 66 , R 70 = independently N or absent; R 11 , R 12 , R 17 , R 21 , R 22 , R 28 , R 31 , R 37 , R 44 , R 55 = independently A, C, U or absent; R1, R9, R 14 , R 15 , R 24 , R 27 , R 34 , R 39 = independently A, G or absent; R7, R 29 , R 32 , R 40 , R 45 = independently A, G, U or absent; R 25 = A, U or absent; R 13 = C, G or absent; R2, R3, R 16 , R 26 , R 30 , R 52 , R 62 , R 64 , R 65 , R 67 , R 68 = independently C, G, U or absent; R 18 , R 35 , R 42 , R 53 , R 59 , R 61 , R 71= independently C, U or absent; R 19 , R 51 = independently G or absent; R 10 , R 20 = independently G, U or absent; R8, R 23 , R 36 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) Including, TREM is conditioned on having one or both of the following properties: no more than 15% of residues are N; or no more than 20 residues are absent.

[0331] In certain embodiments, the TREM disclosed herein has formula III LEU Sequence (SEQ ID NO: 597) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Leu is R0 = Absent; R 38 , R 57 , R 72 = independently A or absent; R 60 = A, C or absent; R4, R5, R 48 , R 50 , R 56 , R 58 , R 69 = independently A, C, G or absent; R6, R 33 , R 43 , R 46 , R 49 , R 63 , R 66 , R 70 = independently N or absent; R 11 , R 12 , R 17 , R 21 , R 22 , R 28 , R 31 , R 37 , R 41 , R 44 , R 55 = independently A, C, U or absent; R1, R9, R 14 , R 15 , R 24 , R 27 , R 34 , R 39 = independently A, G or absent; R7, R 29 , R 32 , R 40 , R 45 = independently A, G, U or absent; R 25 = A, U or absent; R 13 = C, G or absent; R2, R3, R 16 , R 30 , R 52 , R 62 , R 64 , R 67 , R 68 = independently C, G, U or absent; R 18 , R 35 , R 42 , R 53 , R 59 , R 61 , R 65 , R 71 = independently C, U or absent; R 19 , R 51 = independently G or absent; R 10 , R 20 , R 26 = independently G, U or absent; R8, R 23 , R 36 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) Including, TREM is conditioned on having one or both of the following properties: no more than 15% of residues are N; or no more than 20 residues are absent.

[0332] Lysine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I LYS (SEQ ID NO: 598), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Lys is R0 = Absent; R 14 =A or absent; R 40 , R 41 = independently A, C or absent; R 34 , R 43 , R 51 = independently A, C, G or absent; R1, R2, R3, R4, R5, R6, R7, R 11 , R 12 , R 16 , R 21 , R 26 , R 30 , R 31 , R 32 , R 44 , R 45 , R 46 , R 48 , R 49 , R 50 , R 58 , R 62 , R 63 , R 65 , R 66 , R 67 , R 68 , R 69 , R 70 = independently N or absent; R 13 , R 17 , R 59 , R 71 = independently A, C, U or absent; R9, R 15 , R 19 , R 20 , R 25 , R 27 , R 52 , R 56 = independently A, G or absent; R 24 , R 29 , R 72 = independently A, G, U or absent; R 18 , R 57 = independently A, U or absent; R 10 , R 33 = independently C, G, or absent; R 42 , R 61 , R 64 = independently C, G, U or absent; R 28 , R 35 , R36 , R 37 , R 53 , R 55 , R 60 = independently C, U or absent; R8, R 22 , R 23 , R 38 , R 39 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0333] In certain embodiments, the TREM disclosed herein has the formula II LYS (SEQ ID NO: 599), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Lys is R0, R 18 , R 23 = non-existence, R 14 =A or absent; R 40 , R 41 , R 43 = independently A, C or absent; R3, R7 = independently A, C, G or absent; R1, R6, R 11 , R 31 , R 45 , R 48 , R 49 , R 63 , R 65 , R 66 , R 68 = independently N or absent; R2, R 12 , R 13 , R 17 , R 44 , R 67 , R 71 = independently A, C, U or absent; R9, R 15 , R 19 , R 20 , R 25 , R 27 , R 34 , R 50 , R 52 , R 56 , R 70 , R 72 = independently A, G or absent; R5, R 24 , R 26 , R 29 , R 32 , R 46 , R 69 = independently A, G, U or absent; R 57 = A, U or absent; R 10 , R 61= independently C, G, or absent; R4, R 16 , R 21 , R 30 , R 58 , R 64 = independently C, G, U or absent; R 28 , R 35 , R 36 , R 37 , R 42 , R 53 , R 55 , R 59 , R 60 , R 62 = independently C, U or absent; R 33 , R 51 = independently G or absent; R8 = G, U or absent; R 22 , R 38 , R 39 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0334] In certain embodiments, the TREM disclosed herein has formula III LYS (SEQ ID NO: 600), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Lys is R0, R 18 , R 23 = non-existence, R9, R 14 , R 34 , R 41 = independently A or absent; R 40 = A, C or absent; R1, R3, R7, R 31 = independently A, C, G or absent; R 48 , R 65 , R 68 = independently N or absent; R2, R 13 , R 17 , R 44 , R 63 , R 66 = independently A, C, U or absent; R5, R 15 , R 19 , R 20 , R 25 , R 27 , R 29 , R 50 , R 52 , R 56 , R 70 , R 72 = independently A, G or absent; R6, R 24 , R 32 , R 49 = independently A, G, U or absent; R 12 , R 26 , R 46 , R 57 = independently A, U or absent; R 11 , R 28 , R 35 , R 43 = independently C or absent; R 10 , R 45 , R 61 = independently C, G, or absent; R4, R 21 , R 64 = independently C, G, U or absent; R 37 , R 53 , R 55 , R 59 , R 60 , R 62 , R 67 , R 71 = independently C, U or absent; R 33, R 51 = independently G or absent; R8, R 30 , R 58 , R 69 = independently G, U or absent; R 16 , R 22 , R 36 , R 38 , R 39 , R 42 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0335] Phenylanine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I PHE (SEQ ID NO: 601), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Phe is R0, R 23 = non-existent; R9, R 14 , R 38 , R 39 , R 57 , R 72 = independently A or absent; R 71 = A, C or absent; R 41 , R 70 = independently A, C, G or absent; R4, R5, R6, R 30 , R 31 , R 32 , R 34 , R 42 , R 44 , R 45 , R 46 , R 48 , R 49 , R 58 , R 62 , R 63 , R 66 , R 67 , R 68 , R 69 = independently N or absent; R 16 , R 61 , R 65 = independently A, C, U or absent; R 15 , R 26 , R 27 , R 29 , R 40 , R 56 = independently A, G or absent; R7, R 51 = independently A, G, U or absent; R 22 , R 24= independently A, U or absent; R 55 , R 60 = independently C or absent; R2, R3, R 21 , R 33 , R 43 , R 50 , R 64 = independently C, G, U or absent; R 11 , R 12 , R 13 , R 17 , R 28 , R 35 , R 36 , R 59 = independently C, U or absent; R 10 , R 19 , R 20 , R 25 , R 37 , R 52 = independently G or absent; R1 = G, U or absent; R8, R 18 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0336] In certain embodiments, the TREM disclosed herein has the formula II PHE (SEQ ID NO: 602), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Phe is R0, R 18 , R 23 = non-existent; R 14 , R 24 , R 38 , R 39 , R 57 , R 72 = independently A or absent; R 46 , R 71 = independently A, C or absent; R4, R 70 = independently A, C, G or absent; R 45 = A, C, U or absent; R6, R7, R 15 , R 26 , R 27 , R 32 , R 34 , R 40 , R 41 , R 56 , R 69 = independently A, G or absent; R 29 = A, G, U or absent; R5, R9, R 67 = independently A, U or absent; R 35 , R 49 , R 55 , R 60 = independently C or absent; R 21 , R 43 , R 62 = independently C, G, or absent; R2, R 33 , R 68 = independently C, G, U or absent; R3, R 11 , R 12 , R 13 , R 28 , R 30 , R 36 , R 42 , R 44 , R 48 , R 58 , R 59 , R 61 , R 66 = independently C, U or absent; R 10 , R 19 , R 20 , R 25 , R 37 , R 51 , R 52 , R 63 , R64 = independently G or absent; R1, R 31 , R 50 = independently G, U or absent; R8, R 16 , R 17 , R 22 , R 53 , R 54 , R 65 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0337] In certain embodiments, the TREM disclosed herein has formula III PHE (SEQ ID NO: 603), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Phe is R0, R 18 , R 22 , R 23 = non-existent; R5, R7, R 14 , R 24 , R 26 , R 32 , R 34 , R 38 , R 39 , R 41 , R 57 , R 72 = independently A or absent; R 46 = A, C or absent; R 70 = A, C, G or absent; R4, R6, R 15 , R 56 , R 69 = independently A, G or absent; R9, R 45 = independently A, U or absent; R2, R 11 , R 13 , R 35 , R 43 , R 49 , R 55 , R 60 , R 68 , R 71 = independently C or absent; R 33 = C, G or absent; R3, R 28 , R 36 , R 48 , R 58 , R 59 , R 61 = independently C, U or absent; R1, R 10 , R 19 , R 20 , R 21 , R 25 , R 27 , R29 , R 37 , R 40 , R 51 , R 52 , R 63 , R 64 = independently G or absent; R8, R 12 , R 16 , R 17 , R 30 , R 31 , R 42 , R 44 , R 50 , R 53 , R 54 , R 65 , R 66 , R 67 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0338] Proline TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I PRO (SEQ ID NO: 604), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Pro is R0 = non-existence R 14 , R 57 = independently A or absent; R 70 , R 72 = independently A, C or absent; R9, R 26 , R 27 = independently A, C, G or absent; R4, R5, R6, R 16 , R 21 , R 29 , R 30 , R 31 , R 32 , R 33 , R 34 , R 37 , R 41 , R 42 , R 43 , R 44 , R 45 , R 46 , R 48 , R 49 , R 50 , R 58 , R 61 , R 62 , R 63 , R 64 , R 66 , R 67 , R 68 = independently N or absent; R 35 , R 65 = independently A, C, U or absent; R 24 , R 40 , R 56 = independently A, G or absent; R7, R 25 , R 51 = independently A, G, U or absent; R 55 , R 60 = independently C or absent; R1, R3, R 71 = independently C, G, or absent; R 11 , R 12 , R 20 , R 69 = independently C, G, U or absent; R 13 , R 17 , R 18 , R 22 , R 23 , R 28 , R 59= independently C, U or absent; R 10 , R 15 , R 19 , R 38 , R 39 , R 52 , = independently G or absent; R2 = independently G, U, or absent; R8, R 36 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0339] In certain embodiments, the TREM disclosed herein has the formula II PRO (SEQ ID NO: 605), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66-R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Pro is R0, R 17 , R 18 , R 22 , R 23 = non-existent; R 14 , R 45 , R 56 , R 57 , R 58 , R 65 , R 68 = independently A or absent; R 61 = A, C, G or absent; R 43 =N or absent; R 37 = A, C, U or absent; R 24 , R 27 , R 33 , R 40 , R 44 , R 63 = independently A, G or absent; R3, R 12 , R 30 , R 32 , R 48 , R 55 , R 60 , R 70 , R 71 , R 72 = independently C or absent; R5, R 34 , R 42 , R 66 = independently C, G, or absent; R 20 = C, G, U or absent; R 35 , R 41 , R 49 , R 62 = independently C, U or absent; R1, R2, R6, R9, R10 , R 15 , R 19 , R 26 , R 38 , R 39 , R 46 , R 50 , R 51 , R 52 , R 64 , R 67 , R 69 = independently G or absent; R 11 , R 16 = independently G, U or absent; R4, R7, R8, R 13 , R 21 , R 25 , R 28 , R 29 , R 31 , R 36 , R 53 , R 54 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0340] In certain embodiments, the TREM disclosed herein has formula III PRO (SEQ ID NO: 606), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21-R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Pro is R0, R 17 , R 18 , R 22 , R 23 = non-existent; R 14 , R 45 , R 56 , R 57 , R 58 , R65 , R 68 = independently A or absent; R 37 = A, C, U or absent; R 24 , R 27 , R 40 = independently A, G or absent; R3, R5, R 12 , R 30 , R 32 , R 48 , R 49 , R 55 , R 60 , R 61 , R 62 , R 66 , R 70 , R 71 , R 72 = independently C or absent; R 34 , R 42 = independently C, G, or absent; R 43 = C, G, U or absent; R 41 = C, U or absent; R1, R2, R6, R9, R 10 , R 15 , R 19 , R 20 , R 26 , R 33 , R 38 , R 39 , R 44 , R 46 , R 50 , R 51 , R 52 , R 63 , R 64 , R 67 , R 69 = independently G or absent; R 16 = G, U or absent; R4, R7, R8, R 11 , R 13 , R 21 , R 25 , R 28 , R 29 , R 31 , R 35, R 36 , R 53 , R 54 , R 59 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0341] Serine TREM consensus sequence In certain embodiments, the TREM disclosed herein has formula I SER (SEQ ID NO: 607), R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10-R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Ser is R0 = absent; R 14 , R 24 , R 57 = independently A or absent; R 41 = A, C or absent; R2, R3, R4, R5, R6, R7, R9, R 10 , R 11 , R 12 , R 13 , R 16 , R 21 , R 25 , R 26 , R 27 , R 28 , R 30 , R 31 , R 32 , R 33 , R 34 , R 37 , R 42 , R 43 , R 44 , R 45 , R 46 , R 48 , R 49 , R 50 , R 62 , R 63 , R 64 , R 65 , R 66 , R 67 , R 68 , R 69 , R 70 = independently N or absent; R 18 = A, C, U or absent; R 15 , R 40 , R 51 , R 56 = independently A, G or absent; R1, R 29 , R 58 , R 72 = independently A, G, U or absent; R 39 = A, U or absent; R 60 = C or absent; R 38= C, G or absent; R 17 , R 22 , R 23 , R 71 = independently C, G, U or absent; R8, R 35 , R 36 , R 55 , R 59 , R 61 = independently C, U or absent; R 19 , R 20 = independently G or absent; R 52 = G, U or absent; R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) with the proviso that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0342] In certain embodiments, the TREM disclosed herein has the formula II SER (SEQ ID NO: 608) R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 -R 43 -R 44 -R 45 -R 46 -[R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 where R is a ribonucleotide residue and the consensus for Ser is R0, R 23 = non-existence, R 14 , R 24 , R 41 , R 57 = independently A or absent; R 44 = A, C or absent; R 25 , R 45 , R 48 = independently A, C, G or absent; R2, R3, R4, R5, R 37 , R 50 , R 62 , R 66 , R 67 , R 69 , R 70 = independently N or absent; R 12 , R 28 , R 65 = independently A, C, U or absent; R9, R 15 , R 29 , R 34 , R 40 , R 56 , R 63 = independently A, G or absent; R7, R 26 , R 30 , R 33 , R 46 , R 58 , R 72 = independently A, G, U or absent; R 39 = A, U or absent; R 11 , R 35 , R 60 , R 61 = independently C or absent; R 13 , R 38 = independently C, G, or absent; R6, R 17 , R 31 , R 43 , R 64 , R 68 = independently C, G, U or absent; R 36 , R 42 , R 49 , R 55 , R 59 , R 71 = independently C, U or absent; R 10 , R 19 , R 20, R 27 , R 51 = independently G or absent; R1, R 16 , R 32 , R 52 = independently G, U or absent; R8, R 18 , R 21 , R 22 , R 53 , R 54 = independently U or absent; [R 47 ] x =N or absent; For example, x=1~271 (for example, x=1~250, x=1~225, x=1~200, x=1~175, x=1~150, x=1~125, x=1~100, x=1 ~75, x=1~50, x=1~40, x=1~30, x=1~29, x=1~28, x=1~27, x=1~26, x=1~25, x=1~24, x=1~23, x =1~22, x=1~21, x=1~20, x=1~19, x=1~18, x=1~17, x=1~16, x=1~15, x=1~14, x=1~13, x=1~12 , x=1~11, x=1~10, x=10~271, x=20~271, x=30~271, x=40~271, x=50~271, x=60~271, x=70~27 1, x=80~271, x=100~271, x=125~271, x=150~271, x=175~271, x=200~271, x=225~271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=1 8, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271) wi...

Claims

1. (i) A sequence of formula A: [L1] ｙ - [AST domain 1] ｘ -[L2] ｘ - [DH domain] ｘ -[L3] ｘ - [ACH domain] ｘ - [VL domain] ｙ - [TH domain] ｘ -[L4] ｘ - [AST domain 2] ｘ , (A) wherein, independently, [L1] and [VL domain] are optional; each y is independently 0 or 1; each x is 0 or 1; and (ii) an asialoglycoprotein receptor (ASGPR) binding portion A tRNA effector molecule (TREM) comprising: A tRNA effector molecule (TREM), wherein the ASGPR-binding moiety is conjugated to an intranucleotide nucleobase of the TREM, or at one or both termini of the TREM, or within an internucleotide bond of the TREM.

2. 2. The TREM of claim 1 , wherein each x is 1.

3. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding moiety is present in L1.

4. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding portion is present within ASt domain 1.

5. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding moiety is present in L2.

6. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding portion is present within a DH domain.

7. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding portion is present in L3.

8. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding portion is present within the ACH domain.

9. The TREM of any one of claims 1 to 2, wherein the ASGPR binding portion is present within the VL domain.

10. The TREM of any one of claims 1 to 2, wherein the ASGPR binding portion is present within the TH domain.

11. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding portion is present in L4.

12. The TREM of any one of claims 1 to 2, wherein the ASGPR-binding portion is present within ASt domain 2.

13. The TREM according to any one of claims 1 to 12, wherein the nucleobase is a natural nucleobase or a non-natural nucleobase.

14. 14. The TREM of any one of claims 1 to 13, wherein the TREM comprises a sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of a TREM provided in Table 12.

15. 15. The TREM of any one of claims 1 to 14, wherein the TREM comprises at least 5 ribonucleotides (nt), 10nt, 15nt, 20nt, 25nt, 30nt, 35nt, 40nt, 45nt, 50nt, 55nt, 60nt, 65nt, 70nt, or 75nt of a TREM provided in Table 12.

16. 16. The TREM of any one of claims 1-15, wherein the TREM comprises a sequence that differs by no more than 1 ribonucleotide (nt), 2 nt, 3 nt, 4 nt, 5 nt, 6 nt, 7 nt, 8 nt, 9 nt, 10 nt, 12 nt, 14 nt, 16 nt, 18 nt, or 20 nt from a TREM provided in Table 12.

17. The TREM of any one of claims 1 to 16, wherein the TREM is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:622, SEQ ID NO:650, or SEQ ID NO:

653.

18. The ASGPR binding moiety has the structure of formula (III-b): 【Chemistry 85】 or a salt thereof, wherein: Each X is independently O, N(R ７ ), or S; R １ , R ３ , R ４ , and R ５ is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, C(O)-alkyl, C(O)-alkenyl, C(O)-alkynyl, C(O)-heteroalkyl, C(O)-haloalkyl, C(O)-aryl, C(O)-heteroaryl, C(O)-cycloalkyl, or C(O)-heterocyclyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from one or more R ８ optionally substituted with; Or R ３ and R ４ together with the oxygen atom to which they are attached, form one or more R ８ forming a heterocyclyl ring optionally substituted with; R ２ａ is hydrogen or alkyl; R ２ｂ is —C(O)alkyl (e.g., C(O)CH ３ ) and R ６ａ and R ６ｂ each of which is hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, halo, cyano, nitro, -OR Ａ , aryl, heteroaryl, cycloalkyl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl is selected from one or more R ９ optionally substituted with; R ７ is hydrogen, alkyl, or C(O)-alkyl; R ８ and R ９ each is independently hydrogen, halo, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, cycloalkyl, or heterocyclyl; R Ａ is hydrogen, or alkyl, alkenyl, alkynyl; L １ , L ２ , and L ３ each of is independently a linker; each of m, n, and o is independently an integer from 1 to 100; M is a linker, Where: 【Chemistry 86】 A TREM according to any one of claims 1 to 17, wherein represents a point of attachment to a branch point, a further linker, or a TREM, such as a linker, a nucleobase, an internucleotide bond, or a terminus within the TREM sequence.

19. 19. The TREM of claim 18, wherein X is O.

20. R １ , R ３ , R ４ , and R ５ The TREM of any one of claims 18-19, wherein each of is independently hydrogen or alkyl.

21. R ２ａ is hydrogen, R ２ｂ C(O)CH ３ The TREM according to any one of claims 18 to 20,

22. R ６ａ and R ６ｂ The TREM of any one of claims 18 to 21, wherein each of is hydrogen.

23. 23. The TREM of any one of claims 18 to 22, wherein each of m, n, and o is independently an integer from 1 to 10.

24. M, L １ , L ２ , and L ３ 24. The TREM of any one of claims 18-23, wherein each of independently comprises an alkylene, alkenylene, alkynylene, heteroalkylene, or haloalkylene group.

25. M, L １ , L ２ , and L ３ The TREM of any one of claims 18 to 24, wherein each of is independently cleavable or non-cleavable.

26. L １ , L ２ , and L ３ The TREM of any one of claims 18 to 25, wherein each of independently comprises a polyethylene glycol group.

27. 27. The TREM of any one of claims 18-26, wherein M comprises an ester, amide, disulfide, ether, carbonate, aryl, heteroaryl, cycloalkyl, or heterocyclyl group.

28. The ASGPR binding moiety may be galactose (Gal), galactosamine (GalNH ２ 28. The TREM of any one of claims 1 to 27, comprising a Gal, GalNH 2 , or GalNAc moiety, or an N-acetylgalactosamine (GalNAc) moiety or a plurality of Gal, GalNH 2 , or GalNAc moieties.

29. The TREM of any one of claims 1 to 28, wherein the ASGPR binding moiety comprises an ASGPR carbohydrate and an ASGPR linker.

30. The TREM of any one of claims 1 to 29, wherein the ASGPR binding moiety comprises a triantennary GalNAc moiety.

31. The TREM of any one of claims 1 to 30, wherein the TREM is a compound provided in Table 12.

32. 32. The TREM of any one of claims 1 to 31, wherein the TREM retains the ability to support protein synthesis, to be charged by synthetases, to be bound by elongation factors, to introduce amino acids into peptide chains, to support elongation, and / or to support initiation.

33. 33. The TREM of any one of claims 1 to 32, wherein the TREM has a binding affinity for ASGPR of 0.01 nM to 100 mM.

34. The TREM of any one of claims 1 to 33, wherein the TREM further comprises a chemical modification.

35. A pharmaceutical composition comprising the TREM according to any one of claims 1 to 34.

36. 36. The pharmaceutical composition of claim 35, further comprising a pharma- ceutically acceptable ingredient.

37. A lipid nanoparticle formulation comprising the TREM according to any one of claims 1 to 34.

38. A lipid nanoparticle formulation comprising the pharmaceutical composition according to any one of claims 35 to 36.

39. A method of making a TREM according to any one of claims 1 to 34.

40. A composition for use in the treatment of a subject having a disease or disorder associated with PTC, comprising administering to said subject a TREM according to any one of claims 1 to 34, or a pharmaceutical composition according to any one of claims 35 to 36, or a lipid nanoparticle formulation according to any one of claims 37 to 38, thereby treating said subject having said disease or disorder.

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