TREM Compositions and Methods of Use

Abstract

The present invention relates generally to tRNA-based effector molecules having non-natural modifications and methods related thereto.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 255,420, filed October 13, 2021, U.S. Provisional Application No. 63 / 255,426, filed October 13, 2021, U.S. Provisional Application No. 63 / 284,934, filed December 1, 2021, and U.S. Provisional Application No. 63 / 284,946, filed December 1, 2021, the entire contents of each of the foregoing applications being incorporated herein by reference. [Background technology]

[0002] Transfer RNAs (tRNAs) are complex natural RNA molecules with many functions, including protein initiation and elongation. Summary of the Invention [Means for solving the problem]

[0003] The present disclosure features modified tRNA-based effector molecules (TREMs, e.g., TREMs or TREM fragments), and related compositions and uses thereof. As provided herein, TREMs are complex molecules that can mediate a variety of cellular processes. The TREMs disclosed herein contain at least one modification (e.g., a non-natural modification), e.g., on a constituent nucleotide (e.g., a nucleobase or sugar) or in an internucleotide region, e.g., within the TREM backbone. In one aspect, provided herein is a TREM comprising a sequence of Formula A: [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], wherein, independently, [L1] and [VL domain] are optional, and one of [L1], [ASt domain 1], [L2]-[DH domain], [L3], [ACH domain], [VL domain], [TH domain], [L4], and [ASt domain 2] comprises a nucleotide comprising a non-naturally occurring modification.

[0004] In one embodiment, a TREM (a) has the ability to (i) support protein synthesis, (ii) be charged by a synthetase, (iii) be bound by an elongation factor, (iv) introduce an amino acid into a peptide chain, (v) support elongation, or (vi) support initiation; (b) comprise at least X contiguous nucleotides without non-natural modifications, where X is greater than 3, 4, 5, 6, 7, 8, 9, or 10; (c) comprise at least three (but fewer than all) nucleotides of a type (e.g., A, T, C, G, or U) that contain the same non-natural modification; (d) comprise at least X nucleotides of a type (e.g., A, T, C, G, or U) that do not contain non-natural modifications, where X=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; and (e) 1, 2, 3, 4, 5, 6, 7, 8, 9 of a type (e.g., A, T, C, G, or U) that contains a non-natural modification. , 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; and / or (f) a type that does not contain non-naturally occurring modifications (e.g., A, T, C, G, or , 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.

[0005] In one embodiment, the TREM includes feature (a)(i). In one embodiment, the TREM includes feature (a)(ii). In one embodiment, the TREM includes feature (a)(iii). In one embodiment, the TREM includes feature (a)(iv). In one embodiment, the TREM includes feature (a)(v). In one embodiment, the TREM includes feature (a)(vi). In one embodiment, the TREM includes feature (b). In one embodiment, the TREM includes feature (c). In one embodiment, the TREM includes feature (d). In one embodiment, the TREM includes feature (e). In one embodiment, the TREM includes feature (f). In one embodiment, the TREM includes all of features (a)-(f), or a combination thereof.

[0006] In one embodiment, the TREM domain comprising the non-natural modification has a function, eg, a domain function described herein.

[0007] In one aspect, provided herein is a TREM core fragment comprising a 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 , wherein x=1, y=0 or 1, and one of [ASt domain 1], [ACH domain], and [ASt domain 2] contains a nucleotide having a non-natural modification.

[0008] In one embodiment, the TREM has the ability to support protein synthesis. In one embodiment, the TREM has the ability to be charged by a synthetase. In one embodiment, the TREM has the ability to bind to an elongation factor. In one embodiment, the TREM has the ability to introduce an amino acid into a peptide chain. In one embodiment, the TREM has the ability to support elongation. In one embodiment, the TREM has the ability to support initiation.

[0009] In one embodiment, the [ASt domain 1] and / or [ASt domain 2] comprising the non-natural modification has the ability to initiate or elongate a polypeptide chain.

[0010] In one embodiment, the ACH domain containing the non-natural modification is capable of mediating pairing with a codon.

[0011] In one embodiment, y=1 for any one, two, three, four, five, six, or all or a combination of [L1], [L2], [DH domain], [L3], [VL domain], [TH domain], and [L4].

[0012] In one embodiment, y=0 for any one, two, three, four, five, six, or all of [L1], [L2], [DH domain], [L3], [VL domain], [TH domain], or [L4], or a combination thereof.

[0013] In one embodiment, y=1 for the linker [L1], and L1 comprises a nucleotide with a non-natural modification.

[0014] In one embodiment, y=1 for the linker [L2], and L2 comprises a nucleotide with a non-natural modification.

[0015] In one embodiment, y=1 for the [DH domain (DHD)] and the DHD comprises a nucleotide having a non-natural modification. In one embodiment, the DHD comprising the non-natural modification is capable of mediating recognition of an aminoacyl-tRNA synthetase.

[0016] In one embodiment, y=1 for the linker [L3], and L3 comprises a nucleotide with a non-natural modification.

[0017] In one embodiment, y=1 for the [VL domain (VLD)] and the VLD comprises nucleotides with non-natural modifications.

[0018] In one embodiment, y=1 for the [TH domain (THD)] and the THD comprises a nucleotide having a non-natural modification. In one embodiment, the THD comprising the non-natural modification is capable of mediating ribosome recognition.

[0019] In one embodiment, y=1 for the linker [L4] and L4 comprises a nucleotide with a non-natural modification.

[0020] In another aspect, the disclosure provides a TREM fragment comprising a portion of a TREM, wherein 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], the TREM fragment containing non-natural modifications.

[0021] In one embodiment, the TREM fragment comprises one, two, three or all of the following, or any combination thereof: (a) a TREM half (e.g., a 5' half or 3' half derived from a cleavage within the ACH domain, e.g., within the anticodon sequence), (b) a 5' fragment (e.g., a fragment comprising the 5' end derived from a cleavage within the DH domain or ACH domain), (c) a 3' fragment (e.g., a fragment comprising the 3' end derived from a cleavage within the TH domain), or (d) an internal fragment (e.g., derived from a cleavage in any one of the ACH domain, the DH domain, or the TH domain).

[0022] In one embodiment, the TREM fragment comprises (a) a TREM half that comprises a nucleotide with a non-natural modification.

[0023] In one embodiment, the TREM fragment comprises (b) a 5' fragment comprising a nucleotide with a non-natural modification.

[0024] In one embodiment, the TREM fragment comprises (c) a 3' fragment comprising a nucleotide with a non-natural modification.

[0025] In one embodiment, the TREM fragment comprises (d) an internal fragment comprising a nucleotide with a non-natural modification.

[0026] In any of the embodiments of the TREM, TREM core fragment, or TREM fragment disclosed herein, the TREM domain comprises multiple nucleotides, each having a non-natural modification. In one embodiment, the non-natural modification comprises a nucleobase modification, a sugar (e.g., ribose) modification, or a backbone modification. In one embodiment, the non-natural modification is a sugar (e.g., ribose) modification. In one embodiment, the non-natural modification is a 2'-ribose modification, such as a 2'-OMe, a 2'-halo (e.g., 2'-F), a 2'-MOE, or a 2'-deoxy modification. In one embodiment, the non-natural modification is a backbone modification, such as a phosphorothioate modification.

[0027] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the TREM sequence comprises a CCA sequence at an end, e.g., the 3' end. In one embodiment, the TREM sequence does not comprise a CCA sequence at an end, e.g., the 3' end.

[0028] In any embodiment of a TREM, TREM core fragment, or TREM fragment disclosed herein, the non-natural modification is a modification in the base or backbone of the nucleotide, for example, a modification selected from any one of Tables 5, 6, 7, 8, or 9.

[0029] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the non-natural modification is a base modification selected from the modifications listed in Table 5.

[0030] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the non-natural modification is a base modification selected from the modifications listed in Table 6.

[0031] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the non-natural modification is a base modification selected from the modifications listed in Table 7.

[0032] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the non-natural modification is a backbone modification selected from the modifications listed in Table 8.

[0033] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the non-natural modification is a backbone modification selected from the modifications listed in Table 9.

[0034] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the TREM, TREM core fragment, or TREM fragment is encoded by a sequence shown in Table 1, for example, any one of SEQ ID NOs: 1-451.

[0035] In any embodiment of the TREM, TREM core fragment, or TREM fragment disclosed herein, the TREM, TREM core fragment, or TREM fragment is encoded by a consensus sequence selected from any one of SEQ ID NOs: 562-621.

[0036] In embodiments of any of the TREM, TREM core fragment, or TREM fragment disclosed herein, the TREM, TREM core fragment, or TREM fragment is encoded by a sequence shown in Figure 2, e.g., any one of SEQ ID NOs: 622-9757. In one embodiment, the TREM, TREM core fragment, or TREM fragment comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2, e.g., any one of SEQ ID NOs: 622-9757. In one embodiment, the TREM, TREM core fragment, or TREM fragment comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to a TREM shown in Figure 2, e.g., any one of SEQ ID NOs: 622-3284. In one embodiment, the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications (e.g., 2'-ribose modifications or internucleotide modifications, such as 2'OMe, 2'-halo, 2'-MOE, 2'-deoxy, or phosphorothioate modifications) compared to the TREM, TREM core fragment, or TREM fragment shown in Figure 2, e.g., any one of SEQ ID NOs: 622-9757.

[0037] In embodiments of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM, TREM core fragment, or TREM fragment is a TREM shown in Figure 2, e.g., any one of TREM Nos. 1-9757. In one embodiment, the TREM, TREM core fragment, or TREM fragment includes 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications (e.g., 2'-ribose modifications or internucleotide modifications, e.g., 2'OMe, 2'-halo, 2'-MOE, 2'-deoxy, or phosphorothioate modifications) compared to a TREM shown in Figure 2, e.g., any one of TREM Nos. 1-2663.

[0038] In another aspect, the present disclosure provides a pharmaceutical composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein.

[0039] In another aspect, TREM or its related compositions can be used to regulate production parameters (e.g., expression parameters and / or signaling parameters) of RNA corresponding to a nucleic acid sequence that includes an endogenous open reading frame (ORF) with a premature termination codon (PTC), or a polypeptide encoded by that nucleic acid sequence.

[0040] In another aspect, provided herein is a method of modulating a production parameter of an mRNA corresponding to an endogenous open reading frame (ORF) in a subject, or a polypeptide encoded by that ORF, by contacting the subject with a TREM composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein, wherein the TREM, TREM core fragment, or TREM fragment has an anticodon paired with a codon having a first sequence, in an amount and / or for a time sufficient to modulate the production parameter of the mRNA or polypeptide, thereby modulating the production parameter in the subject. In one embodiment, the production parameter comprises a signaling parameter and / or an expression parameter, e.g., as described herein.

[0041] In another aspect, disclosed herein is a method of modulating protein expression in a cell, wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), which ORF comprises a premature termination codon (PTC), the method comprising contacting the cell with a TREM composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein, wherein the TREM, TREM core fragment, or TREM fragment has an anticodon that pairs with the PTC, in an amount and / or for a time sufficient to modulate expression of the encoded protein, thereby modulating expression of the protein in the cell. In one embodiment, the PTC comprises UAA, UGA, or UAG.

[0042] In another aspect, provided herein is a method for increasing protein expression in a subject, wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), the ORF comprising a premature termination codon (PTC), the method comprising contacting the subject with a TREM composition that (i) has an anticodon that pairs with the PTC, (ii) recognizes an aminoacyl-tRNA synthetase specific for Trp, Tyr, Cys, Glu, Lys, Gln, Ser, Leu, Arg, or Gly, (iii) comprises a sequence of Formula A, or (iv) comprises a non-natural modification, in an amount and / or for a time sufficient to increase expression of the protein. In one embodiment, the PTC comprises UAA, UGA, or UAG. In one embodiment, the TREM composition comprises (i). In one embodiment, the TREM composition comprises (ii). In one embodiment, the TREM composition comprises (iii). In one embodiment, the TREM composition comprises (iv). In one embodiment, the TREM composition includes two of (i)-(iv). In one embodiment, the TREM composition includes three of (i)-(iv). In one embodiment, the TREM composition includes each of (i)-(iv).

[0043] In another aspect, the present disclosure provides a method of treating a subject having an endogenous open reading frame (ORF) containing a premature termination codon (PTC), the method comprising providing a TREM composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein (wherein the TREM comprises an anticodon that pairs with the PTC in the ORF), and contacting the subject with the composition comprising the TREM, TREM core fragment, or TREM fragment in an amount and / or for a time sufficient to treat the subject, thereby treating the subject. In one embodiment, the PTC comprises UAA, UGA, or UAG.

[0044] In another aspect, the present disclosure provides a method of treating a subject having a disease or disorder associated with a premature termination codon (PTC), the method comprising providing a TREM composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein, and contacting the subject with the composition comprising a TREM, TREM core fragment, or TREM fragment in an amount and / or for a time sufficient to treat the subject, thereby treating the subject. In one embodiment, the PTC comprises UAA, UGA, or UAG. In one embodiment, the disease or disorder associated with a PTC is a disease or disorder described herein, e.g., cancer or a monogenic disorder.

[0045] In any of the embodiments of the methods disclosed herein, the codon having the first sequence comprises a mutation (e.g., a point mutation, e.g., a nonsense mutation) that results in a premature termination codon (PTC) selected from UAA, UGA, or UAG. In one embodiment, the codon having the first sequence or PTC comprises a UAA mutation. In one embodiment, the codon having the first sequence or PTC comprises a UGA mutation. In one embodiment, the codon having the first sequence or PTC comprises a UAG mutation.

[0046] In another aspect, the disclosure provides a method of making a TREM, TREM core fragment, or TREM fragment disclosed herein, comprising linking a first nucleotide to a second nucleotide to form a TREM.

[0047] In one embodiment, the TREM, TREM core fragment, or TREM fragment is non-naturally occurring (eg, synthetic).

[0048] In one embodiment, the TREM, TREM core fragment, or TREM fragment is produced by cell-free solid phase synthesis.

[0049] In another aspect, the present disclosure provides a method for regulating a tRNA pool in a cell, comprising providing a TREM, TREM core fragment, or TREM fragment disclosed herein and contacting a cell with the TREM, TREM core fragment, or TREM fragment, thereby regulating the tRNA pool in the cell.

[0050] In one aspect, the present disclosure provides a method of contacting a cell, tissue, or subject with a TREM, TREM core fragment, or TREM fragment disclosed herein, comprising contacting the cell, tissue, or subject with a TREM, TREM core fragment, or TREM fragment, thereby contacting the cell, tissue, or subject with the TREM, TREM core fragment, or TREM fragment.

[0051] In another aspect, the present disclosure provides a method of delivering a TREM, a TREM core fragment, or a TREM fragment to a cell, tissue, or subject, comprising providing a cell, tissue, or subject and contacting the cell, tissue, or subject with a TREM, TREM core fragment, or TREM fragment disclosed herein.

[0052] In one aspect, the disclosure provides a method of modulating a tRNA pool in a cell, the method comprising: optionally, obtaining knowledge about the abundance of one or both of (i) and (ii) (e.g., obtaining knowledge about the relative amounts of (i) and (ii) in the cell), where (i) is a tRNA moiety (first tRNA moiety) having an anticodon that pairs with a codon of an ORF having a first sequence, and (ii) is an isoacceptor tRNA moiety (second tRNA moiety) having an anticodon that pairs with a codon other than the codon having the first sequence in the cell; contacting a cell with a TREM, TREM core fragment, or TREM fragment disclosed herein, wherein the TREM, TREM core fragment, or TREM fragment has a codon having a first sequence, or an anticodon that pairs with a codon other than the codon having the first sequence, in an amount and / or for a time sufficient to modulate the relative amounts of the first tRNA moiety and the second tRNA moiety in the cell; This involves regulating the intracellular tRNA pool.

[0053] In another aspect, the disclosure provides a method of modulating a tRNA pool in a subject having an ORF that includes a codon having a first sequence, the method comprising: optionally, obtaining knowledge about the abundance of one or both of (i) and (ii) (e.g., obtaining knowledge about the relative amounts of (i) and (ii) in a subject), where (i) is a tRNA moiety (first tRNA moiety) having an anticodon that pairs with a codon of an ORF having a first sequence, and (ii) is an isoacceptor tRNA moiety (second tRNA moiety) having an anticodon that pairs with a codon other than the codon having the first sequence, in the subject; contacting a subject with a TREM, TREM core fragment, or TREM fragment disclosed herein, wherein the TREM, TREM core fragment, or TREM fragment has a codon having a first sequence, or an anticodon that pairs with a codon other than the codon having the first sequence, in an amount and / or for a time sufficient to adjust the relative amounts of the first tRNA moiety and the second tRNA moiety in the subject; This involves modulating the tRNA pool in the subject.

[0054] In one aspect, the disclosure provides a method of modulating a tRNA pool in a subject having an endogenous ORF that includes a codon that includes a synonymous mutation (synonymous mutation codon, or SMC), the method comprising: Provided is a composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein, wherein the TREM, TREM core fragment, or TREM fragment comprises an isoacceptor tRNA portion that includes an anticodon sequence that pairs with an SMC(TREM); contacting the subject with the composition in an amount and / or for a time sufficient to modulate the tRNA pool in the subject; This involves modulating the tRNA pool in the subject.

[0055] In another aspect, the disclosure provides a method for modulating a tRNA pool in a cell that contains an endogenous ORF that includes a codon that includes SMC, the method comprising: Provided is a composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein, wherein the TREM, TREM core fragment, or TREM fragment comprises an isoacceptor tRNA portion that includes an anticodon sequence that pairs with an SMC(TREM); contacting the cell with a composition comprising a TREM in an amount and / or for a time sufficient to modulate the tRNA pool in the cell; This involves regulating the intracellular tRNA pool.

[0056] In one aspect, the disclosure provides a method of modulating expression of a protein in a cell, where the protein is encoded by a nucleic acid comprising an ORF that includes a codon having a mutation, the method comprising: contacting a cell with a composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein in an amount and / or for a time sufficient to modulate expression of the encoded protein; In this case, the TREM, TREM core fragment, or TREM fragment has an anticodon that pairs with the codon that has the mutation, This involves regulating the expression of proteins within cells.

[0057] In another aspect, the disclosure provides a method of modulating expression of a protein in a subject, where the protein is encoded by a nucleic acid that includes an endogenous ORF, where the ORF includes a codon with a mutation, and the method includes: contacting a subject with a composition comprising a TREM, TREM core fragment, or TREM fragment disclosed herein in an amount and / or for a time sufficient to modulate expression of the encoded protein; In this case, the TREM, TREM core fragment, or TREM fragment has an anticodon that pairs with the codon that has the mutation, This involves modulating the expression of a protein in a subject.

[0058] In one embodiment of any of the methods disclosed herein, the mutation in the ORF is a nonsense mutation that results in a premature stop codon, for example, selected from UAA, UGA, or UAG. In one embodiment, the stop codon is UAA. In one embodiment, the stop codon is UGA. In one embodiment, the stop codon is UAG.

[0059] In one embodiment of any of the methods disclosed herein, the TREM comprises an anticodon that pairs with a stop codon.

[0060] The TREMs of the present disclosure include TREMs, TREM core fragments, and TREM fragments. A TREM, TREM core fragment, or TREM fragment can be modified with a non-naturally occurring modification, for example, to increase the level and / or activity (e.g., stability) of the TREM. For example, a pharmaceutical TREM composition containing a TREM with a non-naturally occurring modification can be administered to a cell, tissue, or subject to regulate their function, for example, in vitro or in vivo. Disclosed herein are TREMs, TREM core fragments, or TREM fragments containing non-naturally occurring modifications, TREM compositions, preparations, methods of making the TREM compositions and preparations, and methods of using them.

[0061] In one embodiment, the TREM, TREM core fragment, and TREM fragment comprises a non-naturally occurring modification that improves the stability or enhances the activity of the TREM, TREM core fragment, or TREM fragment.

[0062] Additional features of any of the above-described 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 features in the enumerated embodiments, drawings, descriptions, examples, or claims.

[0063] 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 following recited embodiments, drawings, descriptions, examples, or claims. [Brief explanation of the drawings]

[0064] [Figure 1] FIG. 1 is a diagram of a representative TREM with a summary of the design guidance (e.g., design guidance 1-6) described herein. [Figure 2-1]

[0023] Figure 1 is a table summarizing exemplary TREMs, TREM core fragments, and TREM fragments described herein. The sequence of each TREM, TREM core fragment, and TREM fragment is provided, with the chemical modification profile annotated as follows: r: ribonucleotide, m: 2'-OMe, *: PS linkage, f: 2'-fluoro, moe: 2'-moe, d: deoxyribonucleotide, 5MeC: 5-methylcytosine. Thus, for example, mA represents 2'-O-methyladenosine, moe5MeC represents a 2'-MOE nucleotide with a 5-methylcytosine nucleobase, and dA represents an adenosine deoxyribonucleotide. The table also provides the results of activity screening described in Examples 6 and 7, along with mass spectrometry characterization of each TREM, TREM core fragment, and TREM fragment. The results of activity screening are presented in columns titled "A" (described in Example 6), "B," "C," and "D" (all three described in Example 7). 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Same as above [Figure 2-144] Same as above [Figure 2-145] Same as above [Figure 2-146] Same as above [Figure 2-147] Same as above [Figure 2-148] Same as above [Figure 2-149] Same as above [Figure 2-150] Same as above [Figure 2-151] Same as above [Figure 2-152] Same as above [Figure 2-153] Same as above [Figure 2-154] Same as above [Figure 2-155] Same as above [Figure 2-156] Same as above [Figure 2-157] Same as above [Figure 2-158] Same as above [Figure 2-159] Same as above [Figure 2-160] Same as above [Figure 2-161] Same as above [Figure 2-162] Same as above [Figure 2-163] Same as above [Figure 2-164] Same as above [Figure 2-165] Same as above [Figure 2-166] Same as above [Figure 2-167] Same as above [Figure 2-168] Same as above [Figure 2-169] Same as above [Figure 2-170] Same as above [Figure 2-171] Same as above [Figure 2-172] Same as above [Figure 2-173] Same as above [Figure 2-174] Same as above [Figure 2-175] Same as above [Figure 2-176] Same as above [Figure 2-177] Same as above [Figure 2-178] Same as above [Figure 2-179] Same as above [Figure 2-180] Same as above [Figure 2-181] Same as above [Figure 2-182] Same as above [Figure 2-183] Same as above [Figure 2-184] Same as above [Figure 2-185] Same as above [Figure 2-186] Same as above [Figure 2-187] Same as above [Figure 2-188] Same as above [Figure 2-189] Same as above [Figure 2-190] Same as above [Figure 2-191] Same as above [Figure 2-192] Same as above [Figure 2-193] Same as above [Figure 2-194] Same as above [Figure 2-195] Same as above [Figure 2-196] Same as above [Figure 2-197] Same as above [Figure 2-198] Same as above [Figure 2-199] Same as above [Figure 2-200] Same as above [Figure 2-201] Same as above [Figure 2-202] Same as above [Figure 2-203] Same as above [Figure 2-204] Same as above [Figure 2-205] Same as above [Figure 2-206] Same as above [Figure 2-207] Same as above [Figure 2-208] Same as above [Figure 2-209] Same as above [Figure 2-210] Same as above [Figure 2-211] Same as above [Figure 2-212] 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[Figure 2-247] Same as above [Figure 2-248] Same as above [Figure 2-249] Same as above [Figure 2-250] Same as above [Figure 2-251] Same as above [Figure 2-252] Same as above [Figure 2-253] Same as above [Figure 2-254] Same as above [Figure 2-255] Same as above [Figure 2-256] Same as above [Figure 2-257] Same as above [Figure 2-258] Same as above [Figure 2-259] Same as above [Figure 2-260] Same as above [Figure 2-261] Same as above [Figure 2-262] Same as above [Figure 2-263] Same as above [Figure 2-264] Same as above [Figure 2-265] Same as above [Figure 2-266] Same as above [Figure 2-267] Same as above [Figure 2-268] Same as above [Figure 2-269] Same as above [Figure 2-270] Same as above [Figure 2-271] Same as above [Figure 2-272] Same as above [Figure 2-273] Same as above [Figure 2-274] Same as above [Figure 2-275] Same as above [Figure 2-276] Same as above [Figure 2-277] Same as above [Figure 2-278] Same as above [Figure 2-279] Same as above [Figure 2-280] Same as above [Figure 2-281] Same as above [Figure 2-282] Same as above [Figure 2-283] Same as above [Figure 2-284] Same as above [Figure 2-285] Same as above [Figure 2-286] Same as above [Figure 2-287] Same as above [Figure 2-288] Same as above [Figure 2-289] Same as above [Figure 2-290] Same as above [Figure 2-291] Same as above [Figure 2-292] Same as above [Figure 2-293] Same as above [Figure 2-294] Same as above [Figure 2-295] Same as above [Figure 2-296] Same as above [Figure 2-297] Same as above [Figure 2-298] Same as above [Figure 2-299] Same as above [Figure 2-300] Same as above [Figure 2-301] Same as above [Figure 2-302] Same as above [Figure 2-303] Same as above [Figure 2-304] Same as above [Figure 2-305] Same as above [Figure 2-306] Same as above [Figure 2-307] Same as above [Figure 2-308] Same as above [Figure 2-309] Same as above [Figure 2-310] Same as above [Figure 2-311] Same as above [Figure 2-312] Same as above [Figure 2-313] Same as above [Figure 2-314] Same as above [Figure 2-315] Same as above 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[Figure 2-385] Same as above [Figure 2-386] Same as above [Figure 2-387] Same as above [Figure 2-388] Same as above [Figure 2-389] Same as above [Figure 2-390] Same as above [Figure 2-391] Same as above [Figure 2-392] Same as above [Figure 2-393] Same as above [Figure 2-394] Same as above [Figure 2-395] Same as above [Figure 2-396] Same as above [Figure 2-397] Same as above [Figure 2-398] Same as above [Figure 2-399] Same as above [Figure 2-400] Same as above [Figure 2-401] Same as above [Figure 2-402] Same as above [Figure 2-403] Same as above [Figure 2-404] Same as above [Figure 2-405] Same as above [Figure 2-406] Same as above [Figure 2-407] Same as above [Figure 2-408] Same as above [Figure 2-409] Same as above [Figure 2-410] Same as above [Figure 2-411] Same as above [Figure 2-412] Same as above [Figure 2-413] Same as above [Figure 2-414] Same as above [Figure 2-415] Same as above [Figure 2-416] Same as above [Figure 2-417] Same as above [Figure 2-418] Same as above [Figure 2-419] Same as above [Figure 2-420] Same as above [Figure 2-421] Same as above [Figure 2-422] Same as above [Figure 2-423] Same as above [Figure 2-424] Same as above [Figure 2-425] Same as above [Figure 2-426] Same as above [Figure 2-427] Same as above [Figure 2-428] Same as above [Figure 2-429] Same as above [Figure 2-430] Same as above [Figure 2-431] Same as above [Figure 2-432] Same as above [Figure 2-433] Same as above [Figure 2-434] Same as above [Figure 2-435] Same as above [Figure 2-436] Same as above [Figure 2-437] Same as above [Figure 2-438] Same as above [Figure 2-439] Same as above [Figure 2-440] Same as above [Figure 2-441] Same as above [Figure 2-442] Same as above [Figure 2-443] Same as above [Figure 2-444] Same as above [Figure 2-445] Same as above [Figure 2-446] Same as above [Figure 2-447] Same as above [Figure 2-448] Same as above [Figure 2-449] Same as above [Figure 2-450] Same as above [Figure 2-451] Same as above [Figure 2-452] Same as above [Figure 2-453] Same as above [Figure 2-454] Same as above [Figure 2-455] Same as above [Figure 2-456] Same as above [Figure 2-457] Same as above [Figure 2-458] Same as above [Figure 2-459] Same as above [Figure 2-460] Same as above [Figure 2-461] Same as above [Figure 2-462] Same as above [Figure 2-463] Same as above [Figure 2-464] Same as above [Figure 2-465] Same as above [Figure 2-466] Same as above [Figure 2-467] Same as above [Figure 2-468] Same as above [Figure 2-469] Same as above [Figure 2-470] Same as above [Figure 2-471] Same as above [Figure 2-472] Same as above [Figure 2-473] Same as above [Figure 2-474] Same as above [Figure 2-475] Same as above [Figure 2-476] Same as above [Figure 2-477] Same as above [Figure 2-478] Same as above [Figure 2-479] Same as above [Figure 2-480] Same as above [Figure 2-481] Same as above [Figure 2-482] Same as above [Figure 2-483] Same as above [Figure 2-484] Same as above [Figure 2-485] Same as above [Figure 2-486] Same as above [Figure 2-487] Same as above [Figure 2-488] Same as above [Figure 2-489] Same as above [Figure 2-490] Same as above [Figure 2-491] Same as above [Figure 2-492] Same as above [Figure 2-493] Same as above [Figure 2-494] Same as above [Figure 2-495] Same as above [Figure 2-496] Same as above [Figure 2-497] Same as above [Figure 2-498] Same as above [Figure 2-499] Same as above [Figure 2-500] Same as above [Figure 2-501] Same as above [Figure 2-502] Same as above [Figure 2-503] Same as above [Figure 2-504] Same as above [Figure 2-505] Same as above [Figure 2-506] Same as above [Figure 2-507] Same as above [Figure 2-508] Same as above [Figure 2-509] Same as above [Figure 2-510] Same as above [Figure 2-511] Same as above [Figure 2-512] Same as above [Figure 2-513] Same as above [Figure 2-514] Same as above [Figure 2-515] Same as above [Figure 2-516] Same as above [Figure 2-517] Same as above [Figure 2-518] Same as above [Figure 2-519] Same as above [Figure 2-520] Same as above [Figure 2-521] Same as above [Figure 2-522] Same as above [Figure 2-523] Same as above [Figure 2-524] Same as above [Figure 2-525] Same as above [Figure 2-526] Same as above [Figure 2-527] Same as above [Figure 2-528] Same as above [Figure 2-529] Same as above [Figure 2-530] Same as above [Figure 2-531] Same as above [Figure 2-532] Same as above [Figure 2-533] Same as above [Figure 2-534] Same as above [Figure 2-535] Same as above [Figure 2-536] Same as above [Figure 2-537] Same as above [Figure 2-538] Same as above [Figure 2-539] Same as above [Figure 2-540] Same as above [Figure 2-541] Same as above [Figure 2-542] Same as above [Figure 2-543] Same as above [Figure 2-544] Same as above [Figure 2-545] Same as above [Figure 2-546] Same as above [Figure 2-547] Same as above [Figure 2-548] Same as above [Figure 2-549] Same as above [Figure 2-550] Same as above [Figure 2-551] Same as above [Figure 2-552] Same as above [Figure 2-553] Same as above [Figure 2-554] Same as above [Figure 2-555] Same as above [Figure 2-556] Same as above [Figure 2-557] Same as above [Figure 2-558] Same as above [Figure 2-559] Same as above [Figure 2-560] Same as above [Figure 2-561] Same as above [Figure 2-562] Same as above [Figure 2-563] Same as above [Figure 2-564] Same as above [Figure 2-565] Same as above [Figure 2-566] Same as above [Figure 2-567] Same as above [Figure 2-568] Same as above [Figure 2-569] Same as above [Figure 2-570] Same as above [Figure 2-571] Same as above [Figure 2-572] Same as above [Figure 2-573] Same as above [Figure 2-574] Same as above [Figure 2-575] Same as above [Figure 2-576] Same as above [Figure 2-577] Same as above [Figure 2-578] Same as above [Figure 2-579] Same as above [Figure 2-580] Same as above [Figure 3] 1 is an image showing the PTC readthrough activity of exemplary TREMs described herein in the four cell lines outlined in both Examples 6 and 7. Activity is shown as a log2 fold change relative to control unmodified TREMs. [Figure 4]1 is an image showing a Western blot analysis of full-length GLA protein rescue in Fabry disease patient fibroblasts and normal healthy fibroblasts upon administration of an exemplary TREM described herein, as described in Example 11. [Figure 5] 1 shows the results of a time course and dose response study in Fabry patient fibroblasts and normal healthy fibroblasts upon administration of an exemplary TREM described herein, as described in Example 11. [Figure 6] 1 is a graph showing the rescue of GLA activity in Fabry patient fibroblasts upon administration of an exemplary TREM described herein, as outlined in Example 11. [Figure 7]

[0023] Figure 1 is a set of graphs showing PTC readthrough and target engagement of TREM in vivo. (A) is a graph showing dose-dependent expression of luciferase from a plasmid in the liver after hydrodynamic delivery. (B) is a graph showing rescue of a luciferase gene harboring a PTC mutation by a plasmid expressing the corresponding TREM. DETAILED DESCRIPTION OF THE INVENTION

[0065] Enumerated Embodiments 1. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2] (I), During the ceremony, independently, the [L1] and [VL domain] are optional; The TREM, wherein a nucleotide within any one of (i) [L1]-[ASt domain 1]-[L2], (ii) [DH domain]-[L3], (iii) [ACH domain], (iv) [VL domain], (v) [TH domain], and [L4]-[ASt domain 2] includes a nucleotide having a non-natural modification.

[0066] 2. The TREM of embodiment 1, wherein the non-natural modification is present on the 2' position of the nucleotide sugar or in the internucleotide region (e.g., a backbone modification).

[0067] 3. The TREM of any one of the preceding embodiments, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), or 2'deoxy modification.

[0068] 4. The TREM of any one of the preceding embodiments, wherein the non-natural modification is a 2'OMe modification.

[0069] 5. The TREM of any one of the preceding embodiments, wherein said non-natural modification is a 2' halo (e.g., 2'F or 2'Cl) modification.

[0070] 6. The TREM of any one of the preceding embodiments, wherein the non-natural modification is a 2'MOE modification.

[0071] 7. The TREM of any one of the preceding embodiments, wherein the non-natural modification is a 2' deoxy modification.

[0072] 8. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present in an internucleotide region (e.g., a backbone modification).

[0073] 9. The TREM of embodiment 8, wherein the non-natural modification is a phosphorothioate modification.

[0074] 11. The TREM of any one of the preceding embodiments, wherein the TREM has a sequence selected from the sequences shown in Figure 2.

[0075] 12. A TREM according to any one of the preceding embodiments, wherein the TREM is the TREM shown in FIG. 2.

[0076] 13. The TREM of any one of the preceding embodiments, wherein the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2.

[0077] 14. The TREM of any one of the preceding embodiments, wherein the TREM comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2.

[0078] 15. The TREM of any one of the preceding embodiments, wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications (e.g., 2'-ribose modifications or internucleotide modifications, such as 2'OMe, 2'-halo, 2'-MOE, 2'-deoxy, or phosphorothiolate modifications) compared to the TREM shown in Figure 2.

[0079] 16. The TREM of any one of the preceding embodiments, wherein the TREM is selected from TREM numbers 1-500, 501-1000, 1001-1500, 1501-2000, 2001-2500, 2501-3000, 3001-3500, 3501-4000, 4001-4500, 4501-5000, 5001-5500, 5501-6000, 6001-6500, 6501-7000, 7001-7500, 7501-8000, 8001-8500, 8501-9000, and 9001-9136 in FIG. 2 .

[0080] 17. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 1 to 500 in Figure 2.

[0081] 18. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 501 to 1000 in Figure 2.

[0082] 19. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 1001 to 2000 in Figure 2.

[0083] 20. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 2001-3000 in Figure 2.

[0084] 21. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 3001 to 4000 in Figure 2.

[0085] 22. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 4001 to 5000 in Figure 2.

[0086] 23. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 5001 to 6000 in Figure 2.

[0087] 24. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 6001 to 7000 in Figure 2.

[0088] 25. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 7001 to 8000 in Figure 2.

[0089] 26. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 8001 to 9000 in Figure 2.

[0090] 27. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of TREM numbers 9001 to 9136 in Figure 2.

[0091] 28. The TREMs are TREM numbers 1 to 100, 101 to 200, 201 to 300, 301 to 400, 401 to 500, 501 to 600, 601 to 700, 701 to 800, 801 to 900, 901 to 1000, 1001 to 1100, 1101 to 1200, 1201 to 1300, 1301 to 1400, 1401 to 1500, 1501 to 1600, 1601 to 1700, 1701 to 1800, 1801 to 1900, 1901 to 2000, 2001 to 2100, 2101 to 2200, 2201 to 2300, 2301 to 2400, 2401 to 2500, 2501 to 2600, 2601 to 2700, 2701 to 2800, 2801 to 2900, 2901 to 3000, 3001 to 3100, 3101 to 3200, 3201 to 3300, 3301 to 3400, 3401 to 3500, 3501 to 3600, 3601 to 3700, 3701 to 3800, 3801 to 3900, 3901 to 4000, 4001 to 4100, 4101 to 4200, 4201 to 4300, 430 2401-2500, 2501-2600, and 2601-2663.

[0092] 29. The TREM of any one of the preceding embodiments, wherein the TREM is selected from SEQ ID NOs: 1-500, 501-1000, 1001-1500, 1501-2000, 2001-2500, 2501-3000, 3001-3500, 3501-4000, 4001-4500, 4501-5000, 5001-5500, 5501-6000, 6001-6500, 6501-7000, 7001-7500, 7501-8000, 8001-8500, 8501-9000, 9001-9500, and 9501-9757.

[0093] 30. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 1-500.

[0094] 31. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 501-1000.

[0095] 32. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 1001-2000.

[0096] 33. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 2001-3000.

[0097] 34. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 3001-4000.

[0098] 35. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 4001-5000.

[0099] 36. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 5001-6000.

[0100] 37. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 6001-7000.

[0101] 38. A TREM according to any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 7001-8000.

[0102] 39. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 8001-9000.

[0103] 40. The TREM of any one of the preceding embodiments, wherein the TREM has at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NOs: 9001-9757.

[0104] 41. The TREM is selected from the group consisting of SEQ ID NOs: 1 to 100, 101 to 200, 201 to 300, 301 to 400, 401 to 500, 501 to 600, 601 to 700, 701 to 800, 801 to 900, 901 to 1000, 1001 to 1100, 1101 to 1200, 1201 to 1300, 1301 to 1400, 1401 to 1500, 1501 to 1600, 1601 to 1700, 1701 to 1800, 1801 to 1900, 2001 to 2100, 2101 to 2200, 2201 to 2300, 2301 to 2400, 2401 to 2500, 2501 to 2600, 2601 to 2700, 2701 to 2800, 2801 to 2900, 2901 to 3000, 3001 to 3100, 3101 to 3200, 3201 to 3300, 3301 to 3400, 3401 to 3500, 3501 to 3600, 3601 to 3700, 3701 to 3800, 3801 to 3900, 3901 to 4000, 4001 to 4100, 4001 to 4200, 4001 to 4300, 4001 to 4 1900, 1901-2000, 2001-2100, 2101-2200, 2201-2300, 2301-2400, 2401-2500, 2501-2600, 2601-2700, 2701-2800, 2801-2900, 2901-3000, 3001-3100, 3101-3200, and 3201-3284.

[0105] 42. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 622.

[0106] 43. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 622; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622.

[0107] 44. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 622; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 622.

[0108] 45. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 622.

[0109] 46. ​​The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0110] 47. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0111] 48. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622 and does not comprise a non-natural modification within the [ASt domain 1].

[0112] 49. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 622.

[0113] 50. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0114] 51. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0115] 52. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622 and does not comprise a non-natural modification within the [DH domain].

[0116] 53. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 622.

[0117] 54. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0118] 55. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0119] 56. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622 and does not comprise a non-natural modification within the [ACH domain].

[0120] 57. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 622.

[0121] 58. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0122] 59. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0123] 60. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622 and does not comprise non-natural modifications within the [VL domain].

[0124] 61. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 622.

[0125] 62. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0126] 63. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0127] 64. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622 and does not comprise a non-natural modification within the [TH domain].

[0128] 65. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 622.

[0129] 66. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0130] 67. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0131] 68. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 622 and does not comprise a non-natural modification within the [ASt domain 2].

[0132] 69. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 623.

[0133] 70. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 623; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 623.

[0134] 71. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 623; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623.

[0135] 72. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 623.

[0136] 73. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0137] 74. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0138] 75. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623 and does not comprise a non-natural modification within the [ASt domain 1].

[0139] 76. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 623.

[0140] 77. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0141] 78. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0142] 79. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623 and does not comprise a non-natural modification within the [DH domain].

[0143] 80. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 623.

[0144] 81. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0145] 82. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0146] 83. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623 and does not comprise a non-natural modification within the [ACH domain].

[0147] 84. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 623.

[0148] 85. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0149] 86. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0150] 87. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623 and does not comprise non-natural modifications within the [VL domain].

[0151] 88. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 623.

[0152] 89. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0153] 90. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0154] 91. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623 and does not comprise a non-natural modification within the [TH domain].

[0155] 92. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 623.

[0156] 93. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0157] 94. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0158] 95. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 623 and does not comprise a non-natural modification within the [ASt domain 2].

[0159] 96. The TREM of any one of the previous embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 624.

[0160] 97. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 624; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 624.

[0161] 98. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 624; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 624.

[0162] 99. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 624.

[0163] 100. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0164] 101. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0165] 102. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624 and does not comprise a non-natural modification within the [ASt domain 1].

[0166] 103. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 624.

[0167] 104. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0168] 105. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0169] 106. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624 and does not comprise a non-natural modification within the [DH domain].

[0170] 107. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 624.

[0171] 108. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0172] 109. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0173] 110. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624 and does not comprise a non-natural modification within the [ACH domain].

[0174] 111. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 624.

[0175] 112. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0176] 113. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0177] 114. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624 and does not comprise non-natural modifications within the [VL domain].

[0178] 115. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 624.

[0179] 116. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0180] 117. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0181] 118. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624 and does not comprise a non-natural modification within the [TH domain].

[0182] 119. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 624.

[0183] 120. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0184] 121. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0185] 122. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 624 and does not comprise a non-natural modification within the [ASt domain 2].

[0186] 123. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6967.

[0187] 124. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6967; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 6967.

[0188] 125. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6967; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 6967.

[0189] 126. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 6967.

[0190] 127. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0191] 128. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0192] 129. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967 and does not comprise a non-natural modification within the [ASt domain 1].

[0193] 130. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 6967.

[0194] 131. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0195] 132. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0196] 133. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967 and does not comprise a non-natural modification within the [DH domain].

[0197] 134. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 6967.

[0198] 135. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0199] 136. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0200] 137. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967 and does not comprise a non-natural modification within the [ACH domain].

[0201] 138. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 6967.

[0202] 139. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0203] 140. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0204] 141. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967 and does not comprise non-natural modifications within the [VL domain].

[0205] 142. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 6967.

[0206] 143. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0207] 144. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0208] 145. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967 and does not comprise a non-natural modification within the [TH domain].

[0209] 146. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 6967.

[0210] 147. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0211] 148. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0212] 149. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6967 and does not comprise a non-natural modification within the [ASt domain 2].

[0213] 150. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4386.

[0214] 151. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4386; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 4386.

[0215] 152. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4386; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 4386.

[0216] 153. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 4386.

[0217] 154. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0218] 155. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0219] 166. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386 and does not comprise a non-natural modification within the [ASt domain 1].

[0220] 167. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 4386.

[0221] 168. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0222] 169. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0223] 170. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386 and does not comprise a non-natural modification within the [DH domain].

[0224] 171. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 4386.

[0225] 172. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0226] 173. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0227] 174. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386 and does not comprise a non-natural modification within the [ACH domain].

[0228] 175. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 4386.

[0229] 176. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0230] 177. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0231] 178. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386 and does not comprise non-natural modifications within the [VL domain].

[0232] 179. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 4386.

[0233] 180. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0234] 181. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0235] 182. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386 and does not comprise a non-natural modification within the [TH domain].

[0236] 183. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 4386.

[0237] 184. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0238] 185. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0239] 186. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386 and does not comprise a non-natural modification within the [ASt domain 2].

[0240] 187. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4834.

[0241] 188. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4834; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 4834.

[0242] 189. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4386; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 4386.

[0243] 190. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 4834.

[0244] 191. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0245] 192. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0246] 193. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834 and does not comprise a non-natural modification within the [ASt domain 1].

[0247] 194. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 4834.

[0248] 195. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0249] 196. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0250] 197. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834 and does not comprise a non-natural modification within the [DH domain].

[0251] 198. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 4834.

[0252] 199. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0253] 200. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0254] 201. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834 and does not comprise a non-natural modification within the [ACH domain].

[0255] 202. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 4834.

[0256] 203. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0257] 204. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0258] 205. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834 and does not comprise non-natural modifications within the [VL domain].

[0259] 206. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 4834.

[0260] 207. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4386, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0261] 208. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0262] 209. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834 and does not comprise a non-natural modification within the [TH domain].

[0263] 210. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 4834.

[0264] 211. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 2].

[0265] 212. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0266] 213. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4834 and does not comprise a non-natural modification within the [ASt domain 2].

[0267] 214. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6749.

[0268] 215. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6749; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 6749.

[0269] 216. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6749; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 6749.

[0270] 217. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 6749.

[0271] 218. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0272] 219. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0273] 220. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749 and does not comprise a non-natural modification within the [ASt domain 1].

[0274] 221. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 6749.

[0275] 222. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0276] 223. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0277] 224. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749 and does not comprise a non-natural modification within the [DH domain].

[0278] 225. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 6749.

[0279] 226. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0280] 227. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0281] 228. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749 and does not comprise a non-natural modification within the [ACH domain].

[0282] 229. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 6749.

[0283] 230. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0284] 231. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0285] 232. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749 and does not comprise non-natural modifications within the [VL domain].

[0286] 233. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 6749.

[0287] 234. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0288] 235. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0289] 236. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749 and does not comprise a non-natural modification within the [TH domain].

[0290] 237. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 6749.

[0291] 238. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 2].

[0292] 239. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0293] 240. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749 and does not comprise a non-natural modification within the [ASt domain 2].

[0294] 241. The TREM of any one of the previous embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 8051.

[0295] 242. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 8051; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 8051.

[0296] 243. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 8051; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 8051.

[0297] 244. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 8051.

[0298] 245. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0299] 246. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0300] 247. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051 and does not comprise a non-natural modification within the [ASt domain 1].

[0301] 248. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6749, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 8051.

[0302] 249. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0303] 250. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0304] 251. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051 and does not comprise a non-natural modification within the [DH domain].

[0305] 252. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 8051.

[0306] 253. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0307] 254. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0308] 255. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051 and does not comprise a non-natural modification within the [ACH domain].

[0309] 256. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 8051.

[0310] 257. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0311] 258. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0312] 259. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051 and does not comprise non-natural modifications within the [VL domain].

[0313] 260. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 8051.

[0314] 261. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0315] 262. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0316] 263. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051 and does not comprise non-natural modifications within the [TH domain].

[0317] 264. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 8051.

[0318] 265. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0319] 266. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0320] 267. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 8051 and does not comprise a non-natural modification within the [ASt domain 2].

[0321] 268. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6707.

[0322] 269. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6707; and / or (ii) The TREM of any one of the preceding embodiments, wherein said TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 6707.

[0323] 270. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6707; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 6707.

[0324] 271. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 6707.

[0325] 272. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0326] 273. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0327] 274. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707 and does not comprise a non-natural modification within the [ASt domain 1].

[0328] 275. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 6707.

[0329] 276. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0330] 277. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0331] 278. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707 and does not comprise a non-natural modification within the [DH domain].

[0332] 279. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 6707.

[0333] 280. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0334] 281. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0335] 282. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707 and does not comprise a non-natural modification within the [ACH domain].

[0336] 283. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 6707.

[0337] 284. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0338] 285. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0339] 286. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707 and does not comprise non-natural modifications within the [VL domain].

[0340] 287. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 6707.

[0341] 288. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0342] 289. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0343] 290. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707 and does not comprise a non-natural modification within the [TH domain].

[0344] 291. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 6707.

[0345] 292. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 2].

[0346] 293. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0347] 294. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 6707 and does not comprise a non-natural modification within the [ASt domain 2].

[0348] 295. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 5630.

[0349] 296. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 5630; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 5630.

[0350] 297. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 5630; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 5630.

[0351] 298. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 5630.

[0352] 299. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0353] 300. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, the non-natural modification is a 2'-O-methyl (2-OMe) modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0354] 301. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630 and does not comprise a non-natural modification within the [ASt domain 1].

[0355] 302. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 5630.

[0356] 303. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0357] 304. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0358] 305. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630 and does not comprise non-natural modifications within the [DH domain].

[0359] 306. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 5630.

[0360] 307. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0361] 308. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0362] 309. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630 and does not comprise non-natural modifications within the [ACH domain].

[0363] 310. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 5630.

[0364] 311. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0365] 312. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0366] 313. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630 and does not comprise non-natural modifications within the [VL domain].

[0367] 314. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 5630.

[0368] 315. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0369] 316. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0370] 317. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630 and does not comprise non-natural modifications within the [TH domain].

[0371] 318. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 5630.

[0372] 319. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 2].

[0373] 320. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0374] 321. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 5630 and does not comprise a non-natural modification within the [ASt domain 2].

[0375] 322. The TREM of any one of the previous embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4249.

[0376] 323. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4249; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 4249.

[0377] 324. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 4249; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 4249.

[0378] 325. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 1] of SEQ ID NO: 4249.

[0379] 326. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0380] 327. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is a 2'-O-methyl (2-OMe) modification, and wherein the non-natural modification is present at a nucleotide position within the [ASt domain 1].

[0381] 328. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249 and does not comprise a non-natural modification within the [ASt domain 1].

[0382] 329. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is present at a nucleotide position within the [DH domain] of SEQ ID NO: 4249.

[0383] 330. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0384] 331. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [DH domain].

[0385] 332. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249 and does not comprise a non-natural modification within the [DH domain].

[0386] 333. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is present at a nucleotide position within the [ACH domain] of SEQ ID NO: 4249.

[0387] 334. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0388] 335. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [ACH domain].

[0389] 336. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249 and does not comprise a non-natural modification within the [ACH domain].

[0390] 337. The TREM of any one of the previous embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is present at a nucleotide position within the [VL domain] of SEQ ID NO: 4249.

[0391] 338. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0392] 339. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [VL domain].

[0393] 340. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249 and does not comprise non-natural modifications within the [VL domain].

[0394] 341. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is present at a nucleotide position within the [TH domain] of SEQ ID NO: 4249.

[0395] 342. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0396] 343. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [TH domain].

[0397] 344. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249 and does not comprise non-natural modifications within the [TH domain].

[0398] 345. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and wherein the non-natural modification is present at a nucleotide position within [ASt domain 2] of SEQ ID NO: 4249.

[0399] 346. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0400] 347. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249, and the non-natural modification is a 2'-O-methyl (2-OMe) or a 2'-halo (e.g., 2'F or 2'Cl) modification, and the non-natural modification is present at a nucleotide position within the [Ast domain 2].

[0401] 348. The TREM of any one of the preceding embodiments, wherein the TREM comprises SEQ ID NO: 4249 and does not comprise a non-natural modification within the [ASt domain 2].

[0402] 349. The TREM is represented by 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 -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 is Ala; R = none; R 14 ,R 57 = A or none independently; R 26 = A, C, G or None; R5, R6, R 15 ,R 16 ,R 21 ,R 30 ,R31 ,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 = N or None independently; R 11 ,R 35 ,R 65 = A, C, U or none independently; R1, R9, R 20 ,R 38 ,R 40 ,R 51 ,R 52 ,R 56 = A, G or none independently; R7, R 22 ,R 25 ,R 27 ,R 29 ,R 46 ,R 53 ,R 72 = independently A, G, U or none; R 24 ,R 69 = independently A, U or none; R 70 ,R 71 = independently C or none; R3, R4 = independently C, G or none; R 12 ,R 33 ,R 36 ,R 62 ,R 68 = independently C, G, U or none; R 13 ,R 17 ,R 28 ,R 39 ,R 55 ,R 60 ,R 61 = independently C, U or none; R 10 ,R 19 ,R 23 = G or None independently; R2 = G, U or None; R8, R 18 ,R 54 = independently U or None; [R47 ] x =N or None, for example, x=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= 70-271,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). 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0403] 350. The TREM of embodiment 349, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0404] 351. The TREM of embodiment 350, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0405] 352. The TREM is represented by 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 -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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Ala is: R0,R 18 = None; R 14 ,R 24 ,R 57 = independently A or None; R 15 ,R 26 ,R 64 = independently A, C, G or none; R 16 ,R 31 ,R 50 ,R 59 = N or None independently; R 11 ,R 32 ,R 37 ,R 41 ,R 43 ,R 45 ,R 49 ,R 65 ,R 66 = independently A, C, U or none; R1, R5, R9, R 25 ,R 27 ,R 38 ,R 40 ,R 46 ,R 51 ,R 56 = independently A, G or none; R7,R 22 ,R 29 ,R 42 ,R 44 ,R 53 ,R 63 ,R 72 = independently A, G, U or none; R6,R 35 ,R 69= independently A, U or None; R 55 ,R 60 ,R 70 ,R 71 = independently C or none; R3 = C, G or None; R 12 ,R 36 ,R 48 = independently C, G, U or none; 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 none; R4,R 10 ,R 19 ,R 20 ,R 23 ,R 52 = independently G or None; R2, R8, R 33 = independently G, U or none; R 21 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0406] 353. The TREM of embodiment 352, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0407] 354. The TREM of embodiment 353, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0408] 355. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Ala is: R0,R 18 = None; R 14 ,R 24 ,R 57 ,R 72 = independently A or None; R 15 ,R 26 ,R 64 = independently A, C, G or none; R 16 ,R 31 ,R 50 = N or None independently; R 11 ,R 32 ,R 37 ,R 41 ,R 43 ,R 45 ,R 49 ,R 65 ,R 66 = independently A, C, U or none; R5, R9, R 25 ,R 27 ,R 38 ,R 40 ,R 46 ,R 51 ,R 56 = independently A, G or none; R7,R 22 ,R 29 ,R 42 ,R 44 ,R 53 ,R 63 = independently A, G, U or none; R6,R 35 = independently A, U or None; R55 ,R 60 ,R 61 ,R 70 ,R 71 = independently C or none; R 12 ,R 48 ,R 59 = independently C, G, U or none; R 13 ,R 17 ,R 28 ,R 30 ,R 34 ,R 39 ,R 58 ,R 62 ,R 67 ,R 68 = independently C, U or none; R1, R2, R3, R4, R 10 ,R 19 ,R 20 ,R 23 ,R 52 = independently G or None; R 33 ,R 36 = independently G, U or none; R8,R 21 ,R 54 ,R 69 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0409] 356. The TREM of embodiment 355, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0410] 357. The TREM of embodiment 356, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0411] 358. The TREM is represented by 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 contains an array of where R is a ribonucleotide residue and the consensus for Arg is: R 57 =A or None; R9,R 27 = independently A, C, G or none; R1, R2, R3, R4, R5, R6, R7, R 11 ,R 12 ,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 = N or None independently; R 13 ,R 17 ,R 41 = independently A, C, U or none; R 19 ,R 20 ,R24 ,R 40 ,R 56 = independently A, G or none; R 14 ,R 15 ,R 72 = independently A, G, U or none; R 18 =A,U or None; R 38 =C or None; R 35 ,R 43 ,R 61 = independently C, G, U or none; R 28 ,R 55 ,R 59 ,R 60 = independently C, U or none; R0,R 10 ,R 52 = independently G or None; R8,R 39 = independently G, U or none; R 36 ,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0412] 359. The TREM of embodiment 358, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0413] 360. The TREM of embodiment 359, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0414] 361. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Arg is: R 18 = None; R 24 ,R 57 = independently A or None; R 41 =A,C or None; R3, R7, R 34 ,R 50 = independently A, C, G or none; R2, R5, R6, R 12 ,R 26 ,R 32 ,R 37 ,R 44 ,R 58 ,R 66 ,R 67 ,R 68 ,R 70 = N or None independently; R 49 ,R 71 = independently A, C, U or none; R1,R 15 ,R 19 ,R 25 ,R 27 ,R 40 ,R 45 ,R 46 ,R 56 ,R 72 = independently A, G or none; R 14 ,R 29 ,R 63 = independently A, G, U or none; R 16 ,R 21 = independently A, U or None; R38 ,R 61 = independently C or none; R 33 ,R 48 = independently C, G or none; R4, R9, R 11 ,R 43 ,R 62 ,R 64 ,R 69 = independently C, G, U or none; R 13 ,R 22 ,R 28 ,R 30 ,R 31 ,R 35 ,R 55 ,R 60 ,R 65 = independently C, U or none; R0,R 10 ,R 20 ,R 23 ,R 51 ,R 52 = independently G or None; R8,R 39 ,R 42 = independently G, U or none; R 17 ,R 36 ,R 53 ,R 54 ,R 59 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0415] 362. The TREM of embodiment 361, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0416] 363. The TREM of embodiment 362, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0417] 364. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Arg is: R 18 = None; R 15 ,R 21 ,R 24 ,R 41 ,R 57 = independently A or None; R 34 ,R 44 = independently A, C or none; R3, R5, R 58 = independently A, C, G or none; R2, R6, R 66 ,R 70 = N or None independently; R 37 ,R 49 = independently A, C, U or none; R1,R 25 ,R 29 ,R 40 ,R 45 ,R 46 ,R 50 = independently A, G or none; R 14 ,R 63 ,R 68 = independently A, G, U or none; R 16 =A,U or None; R 38 ,R 61 = independently C or none; R7,R 11 ,R 12 ,R 26 ,R 48 = independently C, G or none; R64 ,R 67 ,R 69 = independently C, G, U or none; 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 none; R0,R 10 ,R 19 ,R 20 ,R 23 ,R 27 ,R 33 ,R 51 ,R 52 ,R 56 ,R 72 = independently G or None; R8, R9, R 32 ,R 39 ,R 42 = independently G, U or none; R 17 ,R 36 ,R 53 ,R 54 ,R 59 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0418] 365. The TREM of embodiment 364, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0419] 366. The TREM of embodiment 365, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0420] 367. The TREM is represented by formula I ASN (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 -R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71 -R 72 contains an array of where R is a ribonucleotide residue and the consensus for Asn is: R0,R 18 = None; R 41 =A or None; R 14 ,R 48 ,R 56 = independently A, C, G or none; 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 = N or None independently; R 11 ,R 13 ,R 22 ,R 42 ,R 55 ,R 59 = independently A, C, U or none; R9,R 15 ,R 24 ,R 27 ,R 34 ,R 37 ,R 51 ,R 72 = independently A, G or none; R1, R7, R25 ,R 69 = independently A, G, U or none; R 40 ,R 57 = independently A, U or None; R 60 =C or None; R 33 =C,G or None; R 21 ,R 32 ,R 43 ,R 64 = independently C, G, U or none; R3,R 16 ,R 28 ,R 35 ,R 36 ,R 61 = independently C, U or none; R 10 ,R 19 ,R 20 ,R 52 = independently G or None; R 54 =G,U or None; R8,R 23 ,R 38 ,R 39 ,R 53 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0421] 368. The TREM of embodiment 367, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0422] 369. The TREM of embodiment 368, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0423] 370. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Asn is: R0,R 18 = None R 24 ,R 41 ,R 46 ,R 62 = independently A or None; R 59 =A,C or None; R 14 ,R 56 ,R 66 = independently A, C, G or none; R 17 ,R 29 = N or None independently; R 11 ,R 26 ,R 42 ,R 55 = independently A, C, U or none; 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 none; R 44 ,R 45 ,R 58 = independently A, G, U or none; R 40 ,R 57 = independently A, U or None; R5,R 28 ,R60 = independently C or none; R 33 ,R 65 = independently C, G or none; R 21 ,R 43 ,R 71 = independently C, G, U or none; R3, R6, R 13 ,R 22 ,R 32 ,R 35 ,R 36 ,R 61 ,R 63 ,R 64 = independently C, U or none; R7,R 10 ,R 19 ,R 20 ,R 27 ,R 49 ,R 52 = independently G or None; R 54 =G,U or None; R2, R4, R8, R 16 ,R 23 ,R 30 ,R 31 ,R 38 ,R 39 ,R 50 ,R 53 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0424] 371. The TREM of embodiment 370, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0425] 372. The TREM of embodiment 371, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0426] 373. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Asn is: R0,R 18 = None R 24 ,R 40 ,R 41 ,R 46 ,R 62 = independently A or None; R 59 =A,C or None; R 14 ,R 56 ,R 66 = independently A, C, G or none; R 11 ,R 26 ,R 42 ,R 55 = independently A, C, U or none; 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 none; R 44 ,R 45 ,R 58 = independently A, G, U or none; R 57 =A,U or None; R5,R 28 ,R 60 = independently C or none; R 33 ,R 65 = independently C, G or none; R 17 ,R 21 ,R 29 = independently C, G, U or none; R3, R6, R 13 ,R 22 ,R 32 ,R 35 ,R 36 ,R 43 ,R 61 ,R 63 ,R 64 ,R 71 = independently C, U or none; R7,R 10 ,R 19 ,R 20 ,R 25 ,R 27 ,R 49 ,R 52 ,R 72 = independently G or None; R 54 =G,U or None; R2, R4, R8, R 16 ,R 23 ,R 30 ,R 31 ,R 38 ,R 39 ,R 50 ,R 53 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0427] 374. The TREM of embodiment 373, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0428] 375. The TREM of embodiment 374, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0429] 376. The TREM is represented by 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 contains an array of where R is a ribonucleotide residue and the consensus for Asp is: R0=none R 24 ,R 71 = independently A, C or none; R 33 ,R 46 = independently A, C, G or none; 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 = N or None independently; R 13 ,R 21 ,R 34 ,R 41 ,R 57 ,R 65 = independently A, C, U or none; 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 none; R7,R 25 ,R42 = independently A, G, U or none; R 39 =C or None; R 50 ,R 62 = independently C, G or none; R 30 ,R 43 ,R 45 ,R 55 ,R 70 = independently C, G, U or none; R8,R 11 ,R 17 ,R 18 ,R 28 ,R 35 ,R 53 ,R 59 ,R 60 ,R 61 = independently C, U or none; R 19 ,R 52 = independently G or None; R1=G,U or None; R 23 ,R 36 ,R 38 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0430] 377. The TREM of embodiment 376, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0431] 378. The TREM of embodiment 377, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0432] 379. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Asp is: R0,R 17 ,R 18 ,R 23 = independently none; R9,R 40 = independently A or None; R 24 ,R 71 = independently A, C or none; R 67 ,R 68 = independently A, C, G or none; R2, R6, R 66 = N or None independently; R 57 ,R 63 = independently A, C, U or none; 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 none; R7,R 12 ,R 26 ,R 65 = independently A, U or None; R 39 ,R 61 ,R 62 = independently C or none; R3,R 31 ,R 45 ,R 70 = independently C, G or none; R4, R5, R 29 ,R 43 ,R 55 = independently C, G, U or none; R8,R 11 ,R 13 ,R 30 ,R 32 ,R 34 ,R 35 ,R 41 ,R 48 ,R 53 ,R 59 ,R 60 = independently C, U or none; R 15 ,R 19 ,R 20 ,R 25 ,R 42 ,R 50 ,R 52 = independently G or None; R1,R 22 ,R 49 ,R 58 ,R 69 = independently G, U or none; R 16 ,R 21 ,R 28 ,R 36 ,R 38 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0433] 380. The TREM of embodiment 379, wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0434] 381. The TREM of embodiment 380, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0435] 382. The TREM is represented by 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 -R59 -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 contains an array of where R is a ribonucleotide residue and the consensus for Asp is: R0,R 17 ,R 18 ,R 23 = None R9,R 12 ,R 40 ,R 65 ,R 71 = independently A or None; R2,R 24 ,R 57 = independently A, C or none; R6,R 14 ,R 27 ,R 46 ,R 51 ,R 56 ,R 64 ,R 67 ,R 68 = independently A, G or none; R3,R 31 ,R 35 ,R 39 ,R 61 ,R 62 = independently C or none; R 66 =C,G or None; 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 none; R 10 ,R 15 ,R19 ,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 None; R 22 ,R 58 = independently G, U or none; R1, R4, R7, R 11 ,R 13 ,R 16 ,R 21 ,R 26 ,R 28 ,R 36 ,R 38 ,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) 10. The TREM of any one of the preceding embodiments, wherein said TREM has one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0436] 383. The TREM of embodiment 382, ​​wherein the TREM comprises a non-natural modification present at one nucleotide position of [ASt domain 1], [DH domain], [ACH domain], [VL domain], [TH domain], or [ASt domain 2].

[0437] 384. The TREM of embodiment 383, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE), 2'deoxy, or phosphorothioate modification.

[0438] 385. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 701; and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 701; and / or (iii) the TREM comprises the sequence of TREM number 80; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 80 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0439] 386. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 701; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 701.

[0440] 387. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 701.

[0441] 388. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 2951; and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 2951; and / or (iii) the TREM comprises the sequence of TREM number 2330; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 2330 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0442] 388. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 2951; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 2951.

[0443] 389. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 2951.

[0444] 390. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 6047; and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 6047; and / or (iii) the TREM comprises the sequence of TREM number 5426; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 5426 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0445] 391. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6047; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 6047.

[0446] 392. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 6047.

[0447] 393. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 9364, and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 9364; and / or (iii) the TREM comprises the sequence of TREM number 8743; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs in no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications compared to the sequence of TREM number 8743, e.g., non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0448] 394. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 9364; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 9364.

[0449] 395. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 9364.

[0450] 396. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 3795, and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 3795; and / or (iii) the TREM comprises the sequence of TREM number 3174; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 3174 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0451] 397. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 3795; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 3795.

[0452] 398. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 3795.

[0453] 399. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 8524, and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 8524; and / or (iii) the TREM comprises the sequence of TREM number 7903; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM No. 7903 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0454] 400. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 8524; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 8524.

[0455] 401. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 8524.

[0456] 402. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 6725, and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 6725; and / or (iii) the TREM comprises the sequence of TREM number 6104; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 6104 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0457] 403. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 6725; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 6725.

[0458] 404. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 6725.

[0459] 405. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 8712, and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 8712; and / or (iii) the TREM comprises the sequence of TREM number 8091; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 8091 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0460] 406. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 8712; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 8712.

[0461] 407. The TREM of any one of the preceding embodiments, having the sequence of SEQ ID NO: 8712.

[0462] 408. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 9488; and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 9488; and / or (iii) the TREM comprises the sequence of TREM number 8867; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs in no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications compared to the sequence of TREM number 8867, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0463] 409. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 9488; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 9488.

[0464] 410. 9488TREM according to any one of the preceding embodiments, having the sequence of SEQ ID NO: 8712.

[0465] 411. (i) the TREM comprises the nucleotide sequence of SEQ ID NO: 5397; and / or (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 5397; and / or (iii) the TREM comprises the sequence of TREM number 4776; and / or (iv) The TREM of any one of the preceding embodiments, wherein the TREM differs from the sequence of TREM number 4776 by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-natural modifications, such as non-natural modifications selected from 2'-O-methyl (2-OMe), 2'-halo (e.g., 2'F or 2'Cl), 2'-O-methoxyethyl (2'MOE2'deoxy), or phosphorothioate modifications.

[0466] 412. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides of SEQ ID NO: 5397; and / or (ii) The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 5397.

[0467] 413. 9488TREM according to any one of the preceding embodiments, having the sequence of SEQ ID NO: 8712.

[0468] 414. The TREM of any one of the preceding embodiments, wherein the non-natural modification is present at a nucleotide position corresponding to one or more of the nucleotides by the CtNS.

[0469] 415. A pharmaceutical composition comprising a TREM according to any one of embodiments 1 to 414.

[0470] 416. A pharmaceutical composition according to embodiment 415, comprising a pharmaceutically acceptable ingredient, such as an excipient.

[0471] 417. A lipid nanoparticle formulation comprising a TREM according to embodiments 1 to 414.

[0472] 418. A method of making a TREM according to any one of embodiments 1 to 414, comprising linking a first nucleotide to a second nucleotide to form said TREM.

[0473] 419. The method of embodiment 418, wherein the TREM, TREM core fragment, or TREM fragment is synthetic (e.g., non-natural).

[0474] 420. The method of embodiments 418-419, wherein the synthesis is carried out in vitro.

[0475] 421. The method of embodiment 419, wherein the TREM, TREM core fragment, or TREM fragment is produced by cell-free solid-phase synthesis.

[0476] 422. A cell comprising a TREM according to any one of embodiments 1 to 414.

[0477] 423. A cell comprising a TREM, a TREM core fragment, or a TREM fragment produced according to the method described in embodiment 418.

[0478] 424.. A method for modulating a tRNA pool in a cell, comprising: A TREM according to any one of embodiments 1 to 414 is provided, contacting the cells with the TREM; thereby regulating the tRNA pool in the cell.

[0479] 425. A method of contacting a cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414, comprising: contacting the cell, tissue, or subject with the TREM; The contacting method thereby comprises contacting the cell, tissue, or subject with the TREM.

[0480] 426. A method of presenting TREM, comprising: The presentation method comprises contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414, thereby presenting the TREM to the cell, tissue, or subject.

[0481] 427. A method for producing a cell, tissue, or subject contacted with TREM, comprising: contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414; The method of forming the TREM comprises forming a cell, tissue, or subject in contact with the TREM.

[0482] 428. A method of using TREM, comprising: contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414; The method of use includes using the TREM thereby.

[0483] 429. A method of applying TREM to a cell, tissue, or subject, comprising: contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414; The application method thus comprises applying TREM to a cell, tissue, or subject.

[0484] 430. A method of exposing a cell, tissue, or subject to TREM, comprising: contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414; This method of exposure includes exposing a cell, tissue, or subject to TREM.

[0485] 431. A method for forming a mixture of TREM and a cell, tissue, or object, comprising: contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414; The method of forming the TREM comprises forming a mixture of the TREM and the cell, tissue, or subject.

[0486] 432. A method for delivering TREM to a cell, tissue, or subject, comprising: The delivery method comprises providing a cell, tissue, or subject, and contacting the cell, tissue, or subject with a TREM according to any one of embodiments 1 to 414.

[0487] 433. A method for modulating, e.g., ex vivo, metabolism, e.g., translational capacity of an organelle, comprising: The method of modulation comprises providing a preparation of organelles, such as mitochondria or chloroplasts, and contacting said organelles with a TREM according to embodiments 1 to 414.

[0488] 434. A method of treating a subject, e.g., a method of modulating metabolism, e.g., cellular translation capacity, in a subject, comprising: providing, e.g., administering, to the subject a TREM according to embodiments 1 to 414; thereby treating the subject.

[0489] 435. A method for modulating a tRNA pool in a cell, the tRNA pool comprising an endogenous open reading frame (ORF), the ORF comprising a codon having a first sequence, the method comprising: Optionally, obtaining knowledge about the abundance of one or both of (i) and (ii), e.g., obtaining knowledge about the relative amounts of (i) and (ii) in said cell, wherein (i) is a tRNA moiety (first tRNA moiety) having an anticodon that pairs with said codon of said ORF having a first sequence, and (ii) is an isoacceptor tRNA moiety (second tRNA moiety) having an anticodon that pairs with a codon other than said codon having said first sequence in said cell, and wherein said method also comprises: contacting the cell with a TREM of any one of embodiments 1-414, wherein the TREM has an anticodon that pairs with the codon having the first sequence, or with the codon other than the codon having the first sequence, in an amount and / or for a time sufficient to regulate the relative amounts of the first tRNA moiety and the second tRNA moiety in the cell, and wherein the method also thereby regulating the tRNA pool in the cell.

[0490] 436. A method for modulating a tRNA pool in a subject having an endogenous open reading frame (ORF), the ORF including a codon having a first sequence, the method comprising: Optionally, obtaining knowledge about the abundance of one or both of (i) and (ii), e.g., obtaining knowledge about the relative amounts of (i) and (ii) in said subject, wherein (i) is a tRNA moiety (first tRNA moiety) having an anticodon that pairs with said codon of said ORF having a first sequence, and (ii) is an isoacceptor tRNA moiety (second tRNA moiety) having an anticodon that pairs with a codon other than said codon having said first sequence, in said subject, and wherein said method also comprises: contacting the subject with a TREM of any one of embodiments 1-414, wherein the TREM has an anticodon that pairs with the codon having the first sequence, or with the codon other than the codon having the first sequence, in an amount and / or for a time sufficient to adjust the relative amounts of the first tRNA moiety and the second tRNA moiety in the subject, and wherein the method also thereby regulating the tRNA pool in the subject.

[0491] 437. A method for modulating a tRNA pool in a subject having an endogenous open reading frame (ORF) that contains a codon containing a synonymous mutation (synonymous mutation codon, i.e., SMC), the method comprising: 415. The method of claim 1, further comprising providing a composition comprising a TREM according to any one of embodiments 1 to 414, wherein the TREM comprises an isoacceptor tRNA moiety comprising an anticodon sequence that pairs with the SMC (the TREM), and wherein the method also comprises: contacting the subject with the composition in an amount and / or for a time sufficient to modulate the tRNA pool in the subject; thereby regulating the tRNA pool in the subject.

[0492] 438. A method for modulating a tRNA pool in a cell containing an endogenous open reading frame (ORF) that contains a codon containing a synonymous mutation (synonymous mutation codon, or SMC), the method comprising: 415. The method of claim 1, further comprising providing a composition comprising a TREM according to any one of embodiments 1 to 414, wherein the TREM comprises an isoacceptor tRNA moiety comprising an anticodon sequence that pairs with the SMC (the TREM), and wherein the method also comprises: contacting the cell with the composition comprising a TREM in an amount and / or for a time sufficient to modulate the tRNA pool in the cell; thereby regulating the tRNA pool in the cell.

[0493] 439. A method for modulating the expression of a protein in a cell, wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), the ORF comprising a codon having a mutation, the method comprising: contacting the cells with a composition comprising a TREM according to any one of embodiments 1 to 414 in an amount and / or for a time sufficient to modulate expression of the encoded protein; In that case, the TREM has an anticodon that pairs with the codon that has the mutation, and the method also comprises: thereby regulating the expression of the protein in the cell.

[0494] 440. A method for modulating expression of a protein in a subject, wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), the ORF comprising a codon having a mutation, the method comprising: contacting the subject with a composition comprising a TREM according to any one of embodiments 1 to 414 in an amount and / or for a time sufficient to modulate expression of the encoded protein; In that case, the TREM has an anticodon that pairs with the codon that has the mutation, and the method also comprises: thereby regulating expression of the protein in the subject.

[0495] 441. The method of embodiments 439-440, wherein the mutation in the ORF is a nonsense mutation resulting in a premature stop codon, for example selected from UAA, UGA, or UAG.

[0496] 442. The method of embodiments 439-441, wherein the TREM comprises an anticodon that pairs with a stop codon.

[0497] 443. The method of any one of the preceding embodiments, wherein the TREM comprises an anticodon that pairs with a stop codon.

[0498] Other features, objects, and advantages of the invention will become apparent from the description and claims.

[0499] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0500] The present disclosure features tRNA-based effector molecules (TREMs) containing unnatural modifications and related methods. As disclosed herein, TREMs are complex molecules that can mediate various cellular processes. Pharmaceutical TREM compositions, such as TREMs containing unnatural modifications, can be administered to cells, tissues, or subjects to modulate their function. Also disclosed herein are methods for modulating the expression of a protein in a subject or cell (wherein the protein is encoded by a nucleic acid containing a first sequence, e.g., an endogenous open reading frame (ORF) with a mutation, e.g., a premature termination codon (PTC)), and methods for treating a subject with an endogenous open reading frame (ORF) containing a premature termination codon (PTC). Further disclosed herein are TREMs containing unnatural modifications, methods for their production, and compositions containing them.

[0501] definition The term "obtain" or "obtaining," as used herein, refers to obtaining a value, e.g., a numerical value, by "directly obtaining" or "indirectly obtaining" a physical entity or value. "Directly obtaining" refers to performing a process (e.g., performing an analytical method) to obtain a value. "Indirectly obtaining" refers to receiving a value from another entity or source (e.g., a third-party laboratory that directly obtained the value).

[0502] "Diseases or disorders associated with PTC," as the term is used herein, include, but are not limited to, diseases or disorders in which cells express, or at one time express, a polypeptide encoded by an ORF comprising PTC. In some embodiments, the disease associated with PTC is selected from a proliferative disorder (e.g., cancer), a genetic disorder, a metabolic disorder, an immune disorder, an inflammatory disorder, or a neurological disorder. Exemplary diseases or disorders associated with PTC are provided in any one of Tables 15, 16, and 17. In one embodiment, the disease associated with PTC is cancer. In one embodiment, the disease associated with PTC is a monogenic disease.

[0503] The term "isoacceptor," as used herein, refers to multiple tRNA molecules or TREMs, each molecule of the plurality containing a different natural anticodon sequence, and each molecule of the plurality mediating the incorporation of the same amino acid, which is the amino acid that naturally corresponds to the anticodon.

[0504] The term "modification," as used herein with respect to a nucleotide, refers to a modification of the chemical structure of the subject nucleotide, e.g., a covalent modification. The modification can be natural or non-natural. In one embodiment, the modification is non-natural. In one embodiment, the modification is natural. In one embodiment, the modification is a synthetic modification. In one embodiment, the modification is a modification shown in Table 5, 6, 7, 8, or 9.

[0505] "Natural nucleotide," as the term is used herein, refers to a nucleotide that does not contain a non-natural modification. In one embodiment, it contains a natural modification.

[0506] "Non-naturally occurring modification," as the term is used herein with respect to nucleotides, refers to (a) a modification that a cell, e.g., a human cell, does not make to an endogenous tRNA, or (b) a modification that a cell, e.g., a human cell, can make on an endogenous tRNA, but at a position where such modification does not occur on a naturally occurring tRNA, e.g., the modification is within a domain, linker, or arm, or on the nucleotide and / or at a position within a domain, linker, or arm that is essentially free of such modification. In either case, the modification is added synthetically, e.g., in a cell-free reaction, e.g., a solid-phase or solution-phase synthesis reaction. In one embodiment, a non-naturally occurring modification is one that is not present (in identity, site, or position) when the sequence of a TREM is expressed in a mammalian cell, e.g., a HEK293 cell line. Exemplary non-naturally occurring modifications are shown in Tables 5, 6, 7, 8, or 9.

[0507] "Non-naturally occurring modified nucleotide," as that term is used herein, refers to a nucleotide that includes a non-naturally occurring modification at or of the sugar, nucleobase, or phosphate moiety.

[0508] "Nucleotide," as the term is used herein, refers to an entity that includes a sugar, typically a pentameric sugar, a nucleobase, and a phosphate linking group. In one embodiment, 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.

[0509] A "premature stop codon" or "PTC," as the term is used herein, refers to a stop codon occurring within a DNA or mRNA open reading frame (ORF). In one embodiment, a PTC occurs at a position upstream of the natural stop codon within an ORF. In one embodiment, a PTC occurring upstream of a natural stop codon, for example, in an ORF, results in modulation of the production parameters of the corresponding mRNA or polypeptide encoded by the ORF. In one embodiment, a PTC can differ from (or arise from) a pre-mutated sequence due to a point mutation, e.g., a nonsense mutation. In one embodiment, a PTC can differ from (or arise from) a pre-mutated sequence due to a genetic change other than a point mutation, e.g., a genetic abnormality, e.g., a frameshift, deletion, insertion, rearrangement, inversion, translocation, duplication, or transversion. In one embodiment, a PTC results in the production of a truncated protein that lacks natural activity or is associated with a mutation, disease, or other undesirable phenotype. In one embodiment, an ORF containing a PTC is an ORF derived from a tumor suppressor gene. In one embodiment, the mutation that gives rise to a PTC is a driver mutation, eg, a mutation that confers a growth advantage to tumor cells.

[0510] "Production parameters" refer to expression parameters and / or signaling parameters. In one embodiment, the production parameters are expression parameters. Expression parameters include expression parameters of a polypeptide or protein encoded by an endogenous ORF having a first sequence or PTC, or expression parameters of an RNA, e.g., a messenger RNA, encoded by an endogenous ORF having a first sequence or PTC. In one embodiment, the expression parameters are (a) Protein translation, (b) expression levels (e.g., of a polypeptide or protein, or of mRNA); (c) post-translational modifications of polypeptides or proteins; (d) folding (e.g., of a polypeptide or protein, or of mRNA); (e) structure (e.g., of a polypeptide or protein, or of an mRNA); (f) transduction (e.g., of polypeptides or proteins); (g) compartmentalization (e.g., of polypeptides or proteins, or mRNAs); (h) incorporation (e.g., of a polypeptide or protein, or mRNA) into a supramolecular structure, e.g., into a membrane, proteasome, or ribosome; (i) incorporation into multimeric polypeptides, e.g., homo- or heterodimers, and / or (j) may include stability.

[0511] In one embodiment, the generation parameter is a signaling parameter. (1) modulation of a signal transduction pathway, e.g., a cell signal transduction pathway downstream or upstream of a protein encoded by an endogenous ORF having a first sequence or PTC; (2) Regulation of cell fate, (3) Regulation of ribosome occupancy; (4) Regulation of protein translation; (5) Regulation of mRNA stability; (6) Regulation of protein folding and structure, (7) modulation of protein transduction or compartmentalization, and / or (8) It may involve modulation of protein stability.

[0512] "ORF with PTC," as the phrase is used herein, refers to an open reading frame (ORF) that contains a premature termination codon (PTC). In one embodiment, the ORF with PTC is associated with a disease or disorder associated with PTC, e.g., as described herein, e.g., a disease or disorder listed in any one of Tables 15, 16, and 17. In one embodiment, the ORF with PTC is not associated with a disease or disorder associated with PTC.

[0513] "Stop codon," as the term is used herein, refers to a consecutive sequence of three nucleotides in messenger RNA that specifies the end of translation. For example, UAG, UAA, UGA (in RNA) and TAG, TAA, or TGA (in DNA) are stop codons. Stop codons are also known as amber (UAG), ochre (UAA), and opal (UGA).

[0514] "tRNA-based effector molecule" or "TREM," as that term is used herein, refers to an RNA molecule comprising the following structures or characteristics (a)-(v), which may be a recombinant TREM, a synthetic TREM, or a TREM expressed from a heterologous cell. The TREMs described herein are synthetic molecules, e.g., produced in a cell-free reaction, e.g., a solid-phase synthesis reaction or a solution-phase synthesis reaction. The TREMs are chemically distinct from endogenous tRNA molecules made in cells, e.g., mammalian cells, e.g., human cells, e.g., with respect to primary sequence, type or location of modification. A TREM may have multiple (e.g., 2, 3, 4, 5, 6, 7, 8, 9) of the structures and functions (a)-(v).

[0515] In one embodiment, the TREM is non-naturally occurring, as assessed by its structure or method of production.

[0516] In one embodiment, 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) an amino acid binding domain, e.g., an acceptor stem domain (AStD), that binds to an amino acid (e.g., an AStD, when present in, e.g., a wild-type tRNA, 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). Typically, an AStD comprises a 3'-terminal adenosine (CCA) for acceptor stem charge, which is part of synthetase recognition. In one embodiment, the 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 one embodiment, the TREM comprises a fragment or analog of an AStD, e.g., an AStD encoded by a nucleic acid in Table 1, which fragment has AStD activity in some embodiments and no AStD activity in other embodiments. (One of skill in the art can determine from the sequences encoded by the nucleic acids of Table 1 the corresponding sequences associated with any of the domains, stems, loops, or other sequence features described herein. For example, one of skill in the art can determine the sequence corresponding to the AStD of the tRNA sequences encoded by the nucleic acids of Table 1).

[0517] In one embodiment, the AStD corresponds to 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.

[0518] In one embodiment, AStD has formula I ZZZ Residues R1-R2-R3-R4-R5-R6-R7 and residues R 65 -R 66 -R 67 -R 68 -R 69 -R 70 -R 71where ZZZ represents any of the 20 amino acids.

[0519] 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 the 20 amino acids.

[0520] In one embodiment, 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 where ZZZ represents any of the 20 amino acids.

[0521] (a'-1) a linker, e.g., the linker 2 region, comprising residues R8-R9 of the consensus sequence provided in the "Consensus Sequence" section; (b) a dihydrouridine hairpin domain (DHD) (the DHD, e.g., when present in an otherwise wild-type tRNA, mediates aminoacyl-tRNA synthetase recognition, e.g., comprises a sufficient RNA sequence to function as an aminoacyl-tRNA synthetase recognition site for amino acid charging of the TREM). In embodiments, the DHD mediates stabilization of the tertiary structure of the TREM. In one embodiment, 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 one embodiment, the TREM comprises a fragment or analog of a DHD, e.g., a DHD encoded by a nucleic acid in Table 1, which fragment has DHD activity in some embodiments and does not have DHD activity in other embodiments.

[0522] In one embodiment, the DHD corresponds to the corresponding sequence of 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 one embodiment, the DHD has formula I 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 wherein ZZZ represents any of the 20 amino acids; In one embodiment, the DHD is represented by 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 wherein ZZZ represents any of the 20 amino acids; In one embodiment, the DHD is represented by 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 -R20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 wherein ZZZ represents any of the 20 amino acids; (b'-1) Residue R of the consensus sequence provided in the "Consensus Sequence" section 29 a linker comprising, for example, a linker 3 region, (c) an anticodon that binds to the respective codon in the mRNA, e.g., an anticodon hairpin domain (ACHD) (e.g., an ACHD that includes 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 one embodiment, 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 one embodiment, the TREM comprises a fragment or analog of an ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1, which fragment has ACHD activity in some embodiments and does not have ACHD activity in other embodiments.

[0523] In one embodiment, the ACHD corresponds to 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 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 -R46 wherein ZZZ represents any of the 20 amino acids; In one embodiment, ACHD is represented by 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 -R 45 -R 46 wherein ZZZ represents any of the 20 amino acids; In one embodiment, ACHD is represented by 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 wherein ZZZ represents any of the 20 amino acids; (d) Variable Loop Domain (VLD) (The VLD comprises an RNA sequence sufficient to mediate aminoacyl-tRNA synthetase recognition, e.g., to function as an aminoacyl-tRNA synthetase recognition site for amino acid charging of the TREM, e.g., when present in an otherwise wild-type tRNA). In embodiments, the VLD mediates stabilization of the tertiary structure of the TREM. In one embodiment, the VLD modulates, e.g., increases, the specificity of the TREM, e.g., for its cognate amino acid, e.g., the VLD modulates the cognate adapter function of the TREM. In one embodiment, 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 one embodiment, the TREM comprises a fragment or analog of a VLD, e.g., a VLD encoded by a nucleic acid in Table 1, which fragment has VLD activity in some embodiments and no VLD activity in other embodiments.

[0524] In one embodiment, the VLD corresponds to the sequence corresponding to the consensus sequence provided in the "Consensus Sequence" section.

[0525] In one embodiment, the VLD is selected from the consensus sequences provided in the "Consensus Sequences" section, with residues -[R 47 ] xwherein x=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=1-1 2,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=70-2 71,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). (e) a thymine hairpin domain (THD) (the THD comprises a sufficient RNA sequence to mediate ribosome recognition, e.g., to act as a ribosome recognition site and form a TREM-ribosome complex during translation, e.g., when present in an otherwise wild-type tRNA). In one embodiment, the THD has at least 75, 80, 85, 85, 90, 95, or 100% identity to a naturally occurring THD, e.g., a THD encoded by a nucleic acid in Table 1. In one embodiment, the TREM comprises a fragment or analog of a THD, e.g., a THD encoded by a nucleic acid in Table 1, which fragment has THD activity in some embodiments and no THD activity in other embodiments.

[0526] In one embodiment, the THD corresponds to 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 one embodiment, 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 64 wherein ZZZ represents any of the 20 amino acids; In one embodiment, THD is represented by 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 wherein ZZZ represents any of the 20 amino acids; In one embodiment, THD is represented by formula III 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 -R61 -R 62 -R 63 -R 64 wherein ZZZ represents any of the 20 amino acids; (e'1) Residue R of the consensus sequence provided in the "Consensus Sequence" section 72 a linker comprising, for example, a linker 4 region, (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 one embodiment, 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 of Table 1. In one embodiment, 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 of Table 1, which fragment has the activity of the stem or loop structure in some embodiments and not the activity of the stem or loop structure in other embodiments.

[0527] (g) tertiary structures, such as L-shaped tertiary structures; (h) Adaptor function, i.e., TREM, mediates the acceptance of an amino acid, e.g., its cognate amino acid, and the transfer of AA, in the initiation or elongation of a polypeptide chain; (i) cognate adaptor function (TREM mediates acceptance and incorporation of the amino acid naturally associated with the anticodon of the TREM (e.g., the cognate amino acid) to initiate or elongate a polypeptide chain); (j) noncognate adaptor function (TREM mediates acceptance and incorporation of amino acids other than those naturally associated with the anticodon of the TREM (e.g., noncognate amino acids) during initiation or elongation of a polypeptide chain); (k) a regulatory function, such as an epigenetic function (e.g., a gene silencing function or a signaling 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) a structure that allows ribosome binding; (m) post-transcriptional modifications, e.g., natural post-transcriptional modifications; (n) a functional property of tRNA, for example, an ability to inhibit any of the properties (h) to (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 signal transduction pathways, e.g., cell signaling pathways. In one embodiment, the TREM comprises a full-length tRNA molecule or a fragment thereof.

[0528] In one embodiment, the TREM comprises the following characteristics: (a) through (e).

[0529] In one embodiment, the TREM comprises the following characteristics: (a) and (c).

[0530] In one embodiment, the TREM comprises the following characteristics: (a), (c), and (h).

[0531] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), and (b).

[0532] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), and (e).

[0533] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (b), and (e).

[0534] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (b), (e), and (g).

[0535] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), and (m).

[0536] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), and (g).

[0537] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), and (b).

[0538] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), and (e).

[0539] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), (g), (b), and (e).

[0540] In one embodiment, the TREM comprises the following characteristics: (a), (c), (h), (m), (g), (b), (e), and (q).

[0541] In one embodiment, the TREM comprises: (i) an amino acid-binding domain that binds to an amino acid (e.g., an AStD described in (a) of the present specification, and (ii) It contains an anticodon (for example, ACHD described in (c) of the present specification) that binds to each codon in the mRNA.

[0542] In one embodiment, the TREM comprises a flexible RNA linker that provides a covalent bond to (i)-(ii).

[0543] In one embodiment, TREM mediates protein translation.

[0544] In one embodiment, 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 one embodiment, 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 and another domain.

[0545] In one embodiment, 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].

[0546] In one embodiment, 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 one embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In one embodiment, 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 one embodiment, the TREM comprises a TREM domain, e.g., a domain described herein that comprises at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identity to, or differs by 1, 2, 3, 4, 5, 10, or 15 or fewer ribonucleotides to, an RNA encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In one embodiment, the TREM comprises a TREM domain, e.g., a domain described herein that comprises an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In one embodiment, the TREM comprises a TREM domain, e.g., a domain described herein that 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, or a fragment or functional fragment of, a DNA sequence listed in Table 1.

[0547] In one embodiment, the TREM is 76 to 90 nucleotides in length. In 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.

[0548] In one embodiment, the TREM is aminoacylated, eg, charged with an amino acid, by an aminoacyl-tRNA synthetase.

[0549] In one embodiment, the TREM is not charged with an amino acid, eg, an uncharged TREM (uTREM).

[0550] In one embodiment, the TREM comprises a less than full-length tRNA. In embodiments, the TREM may correspond to a naturally occurring fragment of the tRNA, or a non-naturally occurring fragment. Exemplary fragments include a TREM half (e.g., a 5' half or 3' half, e.g., derived from a cleavage within the ACHD, e.g., within the anticodon sequence), a 5' fragment (e.g., a fragment comprising the 5' end from a cleavage within the DHD or ACHD), a 3' fragment (e.g., a fragment comprising the 3' end from a cleavage within the THD), or an internal fragment (e.g., derived from a cleavage in one or more of the ACHD, DHD, or THD).

[0551] A "TREM core fragment," as that term is used herein, is a fragment 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.

[0552] As used herein, "TREM fragment" refers to a portion of a 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].

[0553] A "cognate adaptor function TREM," as that term is used herein, refers to a TREM that mediates initiation or elongation by an AA that is naturally associated with the anticodon of the TREM (cognate AA).

[0554] "Decreased expression," as the term is used herein, refers to a decrease compared to a reference, for example, when an alteration of a regulatory region or addition of an agent results in decreased expression of the product of interest, where expression is decreased compared to other similar cells without the alteration or addition.

[0555] "Exogenous nucleic acid," as the term is used herein, refers to a nucleic acid sequence that is not present in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced, or that differs from the closest sequence by at least one nucleotide. In one embodiment, the exogenous nucleic acid comprises a nucleic acid encoding a TREM.

[0556] "Exogenous TREM," as the term is used herein, refers to TREM that: (a) differs from the closest tRNA sequence in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced, by at least one nucleotide or one post-transcriptional modification; (b) whether it is introduced into cells other than the cells in which it was transfected; (c) present in a cell other than a native cell; or (d) having a non-wild-type expression characteristic, e.g., a level or distribution, e.g., expressed at a higher level than wild-type. In one embodiment, the expression characteristic may be mediated by a change introduced into the expression-regulating nucleic acid or by the addition of an agent that regulates expression of the RNA molecule. In one embodiment, the exogenous TREM comprises one, two, three, or four of characteristics (a)-(d).

[0557] "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 one embodiment, a GMP-grade composition can be used as a pharmaceutical product.

[0558] As used herein, the terms "increase" and "decrease" refer to modulation that results in an increase or decrease, respectively, in the amount of function, expression, or activity of a particular metric compared to a reference. For example, after administration of a TREM described herein to a cell, tissue, or subject, the amount of a marker for a metric described herein (e.g., protein translation, mRNA stability, protein folding) may increase or decrease 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 metric may be measured after administration at a time when the recited effect is achieved, for example, at least 12 hours, 24 hours, 1 week, 1 month, 3 months, or 6 months after initiation of treatment.

[0559] "Increased expression," as the term is used herein, refers to an increase compared to a reference, for example, when an alteration of a regulatory region or addition of an agent results in increased expression of a product of interest, where expression is increased compared to other similar cells without the alteration or addition.

[0560] "Non-cognate adapter function TREM," as that term is used herein, refers to a TREM that mediates initiation or extension by an AA (non-cognate AA) other than the AA naturally associated with the anticodon of the TREM. In one embodiment, a non-cognate adapter function TREM is also referred to as a mischarged TREM (mTREM).

[0561] "Non-naturally occurring sequence," as the term is used herein, refers to a sequence in which adenine is replaced by a residue other than an adenine analog, cytosine is replaced by a residue other than a cytosine analog, guanine is replaced by a residue other than a guanine analog, and uracil is replaced by a residue other than a uracil analog. 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.

[0562] "Pharmaceutical TREM composition," as the term is used herein, refers to a TREM composition suitable for pharmaceutical use. Typically, a pharmaceutical TREM composition includes a pharmaceutical excipient. In one embodiment, the TREM is the only active ingredient in the pharmaceutical TREM composition. In embodiments, the pharmaceutical TREM composition is free, substantially free, or contains less than pharmaceutically acceptable amounts of host cell proteins, DNA, e.g., host cell DNA, endotoxins, and bacteria.

[0563] "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 one embodiment, the covalent modification occurs post-transcriptionally. In one embodiment, the covalent modification occurs concomitantly with transcription. In one embodiment, the modification occurs in vivo, e.g., in cells used to produce the TREM. In one embodiment, the modification occurs ex vivo, e.g., on a TREM isolated or obtained from cells that produced the TREM. In one embodiment, the post-transcriptional modification is selected from the post-transcriptional modifications listed in Table 2.

[0564] "Synthetic TREM," as that term is used herein, refers to a TREM synthesized outside of or near a cell that contains an 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.

[0565] "Recombinant TREM," as the term is used herein, refers to a TREM that is expressed in a cell that has been modified by human intervention and has modifications that mediate the production of the TREM, e.g., the cell includes an exogenous sequence encoding the TREM or a modification that mediates expression, e.g., transcriptional expression or post-transcriptional modification of the TREM. A recombinant TREM can have the same or different sequence, set of post-transcriptional modifications, or tertiary structure as a reference tRNA, e.g., a naturally occurring tRNA.

[0566] "tRNA," as the term is used herein, refers to a naturally occurring transfer ribonucleic acid in its natural state.

[0567]

[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 one embodiment, the composition comprises only a single species of TREM, TREM core fragment, or TREM fragment. In one embodiment, the 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 one embodiment, the TREM composition comprises X TREM, TREM core fragment, or TREM fragment species, where X=2, 3, 4, 5, 6, 7, 8, 9, or 10. In one embodiment, the TREM, TREM core fragment, or TREM fragment has at least 70, 75, 80, 85, 90, or 95% identity, or 100% identity, to a sequence encoded by a nucleic acid of Table 1. The TREM composition may comprise one or more TREMs, TREM core fragments, or TREM fragments. In one embodiment, 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 substantially all liquid has been removed, e.g., after lyophilization). In one embodiment, the composition is liquid. In one embodiment, the composition is dry, e.g., a lyophilized material. In one embodiment, the composition is a frozen composition. In one embodiment, the composition is sterile. In one embodiment, 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 determined by dry weight) of TREM.

[0568] In one embodiment, at least X% of the TREMs in the TREM composition have a non-natural modification at a selected position, where X is 80, 90, 95, 96, 97, 98, 99, or 99.5.

[0569] In one embodiment, at least X% of the TREMs in the TREM composition have a non-natural modification at a first position and a non-natural modification at a second position, where X is independently 80, 90, 95, 96, 97, 98, 99, or 99.5. In an embodiment, the modifications at the first and second positions are the same. In an embodiment, the modifications at the first and second positions are different. In an embodiment, the nucleotides at the first and second positions are the same, e.g., both are adenine. In an embodiment, the nucleotides at the first and second positions are different, e.g., one is adenine and the other is thymine.

[0570] In one embodiment, at least X% of the TREMs in the TREM composition have a non-natural modification at a first position and less than Y% have a non-natural 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 an embodiment, the nucleotides at the first and second positions are the same, e.g., both are adenine. In an embodiment, the nucleotides at the first and second positions are different, e.g., one is adenine and the other is thymine.

[0571] TREM, TREM core fragments, and TREM fragments "tRNA-based effector molecule" or "TREM" refers to an RNA molecule that contains one or more of the properties described herein. A TREM may contain non-natural modifications, for example, as shown in Tables 4, 5, 6, or 7.

[0572] In one embodiment, the TREM includes 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 (wherein the TREM comprises the sequence of Formula A).

[0573] In one embodiment, 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]. In one embodiment, [VL domain] is optional. In one embodiment, [L1] is optional.

[0574] 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. In one embodiment, y=0. In one embodiment, y=1.

[0575] In one embodiment, the TREM fragment comprises a portion of TREM, wherein the TREM 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], and the TREM fragment comprises one, two, three, or all of the following, or any combination thereof: a TREM half (e.g., a 5' half or 3' half derived from a cleavage within the ACH domain, e.g., within the anticodon sequence), a 5' fragment (e.g., a fragment comprising the 5' end derived from a cleavage within the DH domain or ACH domain), a 3' fragment (e.g., a fragment comprising the 3' end derived from a cleavage within the TH domain), 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., a 5' TREM half or a 3' TREM half, e.g., derived from a cleavage within the ACHD), a 5' fragment (e.g., a fragment including the 5' end, derived from a cleavage within the DHD or ACHD), a 3' fragment (e.g., a fragment including the 3' end of the TREM, derived from a cleavage within the THD), or an internal fragment (e.g., derived from a cleavage in one or more of the ACHD, DHD, or THD).

[0576] In one embodiment, a TREM, TREM core fragment, or TREM fragment may be charged with an amino acid (e.g., a cognate amino acid), a non-cognate amino acid (e.g., a mischarged TREM (mTREM)), or no amino acid (e.g., an uncharged TREM (uTREM)). In one embodiment, a 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.

[0577] In some embodiments, the non-extended anticodon is an anticodon of three nucleotides or less. In one embodiment, the non-extended codon pairs with three or fewer codon nucleotides on the nucleic acid to be translated.

[0578] In one embodiment, the TREM, TREM core fragment, or TREM fragment is a cognate TREM. In one embodiment, the TREM, TREM core fragment, or TREM fragment is a non-cognate TREM. In one embodiment, the TREM, TREM core fragment, or TREM fragment recognizes a codon shown in Table 2 or Table 3. [Table 2] [Table 3]

[0579] In one embodiment, 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 an embodiment, the TREM comprises an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence shown 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.

[0580] In one embodiment, 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 one embodiment, 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 contiguous 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.

[0581] 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: 1-451 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: 1-451 disclosed in Table 1.

[0582] 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 that is at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a DNA sequence provided in Table 1, for example, any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0583] 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. [Table 1-1]

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

Table 1-8

Table 1-9

Table 1-10

Table 1-11

Table 1-12

Table 1-13

Table 1-14

Table 1-15

Table 1-16

Table 1-17

Table 1-18

[0584] Non-natural modifications The TREM, TREM core fragment, or TREM fragment described herein may comprise a non-naturally occurring modification, such as a modification described in any one of Tables 5-9. Non-naturally occurring modifications may be made according to methods known in the art. Exemplary methods for making non-naturally occurring modifications are provided in Examples 4-7.

[0585] In one embodiment, a non-naturally occurring modification is a modification that a cell, e.g., a human cell, does not make to an endogenous tRNA.

[0586] In one embodiment, the non-natural modification is a modification that a cell, for example, a human cell, can make to an endogenous tRNA, where the modification is at a position that does not occur in natural tRNA. In one embodiment, the non-natural modification is in a domain, linker, or arm that does not naturally have such a modification. In one embodiment, the non-natural modification is in a site within a domain, linker, or arm that does not naturally have such a modification. In one embodiment, the non-natural modification is on a nucleotide that does not naturally have such a modification. In one embodiment, the non-natural modification is on a nucleotide at a site within a domain, linker, or arm that does not naturally have such a modification.

[0587] In one embodiment, a TREM, TREM core fragment, or TREM fragment described herein comprises a non-naturally occurring modification shown in Table 5, or a combination thereof. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7] [Table 5-8] [Table 5-9] [Table 5-10]

[0588] In one embodiment, a TREM, TREM core fragment, or TREM fragment described herein comprises a modification, or a combination thereof, set forth in Table 6. The modifications set forth in Table 6 occur naturally in RNA and are used herein on a synthetic TREM, TREM core fragment, or TREM fragment in a position that does not occur in nature. [Table 6-1] [Table 6-2] [Table 6-3]

[0589] In one embodiment, a TREM, TREM core fragment, or TREM fragment described herein comprises a non-naturally occurring modification shown in Table 7, or a combination thereof. [Table 7-1] [Table 7-2] [Table 7-3]

[0590] In one embodiment, a TREM, TREM core fragment, or TREM fragment described herein comprises a non-naturally occurring modification shown in Table 8, or a combination thereof. [Table 8-1] [Table 8-2]

[0591] In one embodiment, a TREM, TREM core fragment, or TREM fragment described herein comprises a non-naturally occurring modification shown in Table 9, or a combination thereof. [Table 9]

[0592] Design Guidance The present disclosure further describes exemplary design principles for introducing non-natural modifications into a TREM, TREM core fragment, or TREM fragment. Without being bound by theory, these design principles may provide guidance for adjusting the parameters of the TREM, TREM core fragment, or TREM fragment described herein. These design principles are also referred to herein as "design guidance." For example, a TREM containing a non-natural modification pattern according to the design guidance may exhibit improved stability, for example, in vitro or in cells. Exemplary design guidance is described in detail below.

[0593] Design Guidance 1 In one embodiment, the TREM comprises a sequence of formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], wherein, independently, [L1] and [VL domain] are optional, and one of [L1], [ASt domain 1], [L4], and [ASt domain 2] comprises a nucleotide having a non-natural modification.

[0594] (a) In one embodiment, the nucleotide having the non-natural modification is at any one of nucleotide positions 1-6 or 66-76, for example, (i) the nucleotide positions correspond to 1-6 or 66-76 of SEQ ID NO: 622, or (ii) the nucleotide positions correspond to 1-6 or 66-76 according to CtNS.

[0595] (b) In one embodiment, one of [L1], [ASt domain 1], [L4], and [ASt domain 2] comprises a nucleotide having a 2'-O-methoxy (2'O-Me) modification. In one embodiment, the nucleotide having a 2'OMe modification is at any one of nucleotide positions 1-6, 65-70, and 74-76, e.g., (i) nucleotide positions corresponding to 1-6, 65-70, and 74-76 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 1-6, 65-70, and 74-76 of CtNS.

[0596] (c) In one embodiment, one of [L1], [ASt domain 1], [L4], and [ASt domain 2] comprises a nucleotide having a phosphorothioate (PS) modification. In one embodiment, the PS modification is present at any one of nucleotide positions 1-6 or 65-76, e.g., (i) nucleotide positions corresponding to 1-6 or 65-76 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 1-6 or 65-76 according to CtNS.

[0597] (d) In one embodiment, one of [L1], [ASt domain 1], [L4], and [ASt domain 2] comprises a nucleotide having a 2'-O-methoxy (2'O-Me) modification and a nucleotide having a phosphorothioate (PS) modification. In one embodiment, both the 2'OMe modification and the PS modification are independently present at any one of nucleotide positions 1-6 or 65-76, e.g., (i) the nucleotide positions correspond to 1-6 or 65-76 of SEQ ID NO: 622, or (ii) the nucleotide positions correspond to 1-6 or 65-76 according to CtNS.

[0598] (e) In one embodiment, one of [L1], [ASt domain 1], [L4], and [ASt domain 2] comprises nucleotides having a 2'-O-methoxy (2'O-Me) modification and nucleotides having a phosphorothioate (PS) modification, with the proviso that the nucleotides corresponding to positions 71-73 (e.g., corresponding to SEQ ID NO: 622 or CtNS) do not comprise a 2'-O-methoxy (2'O-Me) modification.

[0599] (f) In one embodiment, one of [L1], [ASt domain 1], [L4], and [ASt domain 2] comprises a nucleotide having a phosphorothioate (PS) modification at one of positions 1-3 or 74-76 (e.g., corresponding to SEQ ID NO: 622 or CtNS).

[0600] Design Guidance 2 In one embodiment, the TREM comprises a sequence of formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], where, independently, [L1] and [VL domain] are optional, and one of [DH domain]-[L3] comprises a nucleotide having a non-natural modification.

[0601] (a) In one embodiment, the nucleotide having the non-natural modification is at any one of nucleotide positions 10-13 or 22-25, e.g., (i) nucleotide positions correspond to 10-13 or 22-25 of SEQ ID NO: 622, or (ii) nucleotide positions correspond to 10-13 or 22-25 according to CtNS.

[0602] (b) In one embodiment, one of the [DH domains]-[L3] includes a nucleotide having a 2'-O-methoxy (2'O-Me) modification. In one embodiment, the nucleotide having the 2'OMe modification is at any one of nucleotide positions 10-13 or 22-25, e.g., (i) nucleotide positions corresponding to 10-13 or 22-25 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 10-13 or 22-25 of CtNS.

[0603] (c) In one embodiment, one of the [DH domain]-[L3] comprises a nucleotide without a phosphorothioate (PS) modification. In one embodiment, the nucleotide without a PS modification is at any one of nucleotide positions 10-13 or 22-25, e.g., (i) nucleotide positions corresponding to 10-13 or 22-25 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 10-13 or 22-25 of CtNS.

[0604] Design Guidance 3 In one embodiment, the TREM comprises a sequence of formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], wherein, independently, [L1] and [VL domain] are optional, and [ACH domain] does not comprise a 2'-nucleotide sugar modification (e.g., a 2'-ribose modification).

[0605] (a) In one embodiment, the [ACH domain] comprises a nucleotide having an internucleotide modification. In one embodiment, the nucleotide having the internucleotide modification is present at any one of nucleotide positions 32-28, e.g., (i) the nucleotide position corresponds to any one of 32-38 of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to any one of 32-38 of CtNS.

[0606] (b) In one embodiment, the [ACH domain] comprises a nucleotide having a phosphorothioate (PS) modification. In one embodiment, the nucleotide having the PS modification is present at any one of nucleotide positions 32-28, e.g., (i) the nucleotide position corresponds to any one of 32-38 of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to any one of 32-38 of CtNS.

[0607] (c) In one embodiment, the [ACH domain] comprises nucleotides without a 2'-nucleotide sugar modification. In one embodiment, the [ACH domain] comprises nucleotides without a 2'-O-methoxy (2'O-Me) modification.

[0608] (d) In one embodiment, the [ACH domain] comprises nucleotides having an internucleotide modification (e.g., a PS modification) and nucleotides without a 2'-nucleotide sugar modification (e.g., a 2'O-Me modification).

[0609] (e) In one embodiment, the [ACH domain] comprises a nucleotide having a 2'-fluoro (2'F) modification. In one embodiment, the nucleotide having a 2'F modification is at nucleotide position 33, e.g., (i) the nucleotide position corresponds to 33 in SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 33 according to CtNS.

[0610] (f) In one embodiment, the [ACH domain] comprises (i) a nucleotide having an internucleotide modification (e.g., a PS modification), (ii) a nucleotide having a 2'F modification, and (iii) a nucleotide without a 2'OMe modification. In one embodiment, the nucleotide having a 2'F modification is at nucleotide position 33, e.g., (i) the nucleotide position corresponds to 33 in SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 33 according to CtNS.

[0611] (g) In one embodiment, the [ACH domain] comprises a nucleotide having a phosphorothioate (PS) modification on the anticodon, e.g., at any one of positions 34-36, where (i) the nucleotide position corresponds to any one of positions 34-36 of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to any one of positions 34-36 according to CtNS.

[0612] Design Guidance 4 In one embodiment, the TREM comprises a sequence of formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], where, independently, [L1] and [VL domain] are optional, and [VL domain] comprises nucleotides having a non-natural modification.

[0613] (a) In one embodiment, the [VL domain] comprises a nucleotide having a 2'-nucleotide sugar modification, for example, (i) the nucleotide having a 2'-nucleotide sugar modification corresponds to any one of 44-48 in SEQ ID NO: 622, or (ii) the nucleotide having a 2'-nucleotide sugar modification corresponds to any one of V1-V27 and 46-48 in accordance with CtNS.

[0614] (b) In one embodiment, the [VL domain] comprises a nucleotide having a 2'-OMe modification. In one embodiment, the 2'OMe modification is present at any one of nucleotide positions in the [VL domain], for example, (i) nucleotide position corresponding to any one of 44-48 of SEQ ID NO: 622, or (ii) nucleotide position corresponding to any one of V1-V27 and 46-48 according to CtNS.

[0615] (b) In one embodiment, the [VL domain] comprises a nucleotide having a 2'-F modification. In one embodiment, the F modification is present at any one of nucleotide positions in the [VL domain], for example, (i) nucleotide position corresponding to any one of 44-48 of SEQ ID NO: 622, or (ii) nucleotide position corresponding to any one of V1-V27 and 46-48 according to CtNS.

[0616] (d) In one embodiment, the [VL domain] comprises nucleotides without an internucleotide modification. In one embodiment, the [VL domain] comprises nucleotides without a PS modification, for example, (i) nucleotide positions corresponding to any one of 44-48 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to any one of V1-V27 and 46-48 according to CtNS.

[0617] (e) In one embodiment, the [VL domain] comprises (i) a nucleotide having a 2'-OMe modification, (ii) a nucleotide having a 2'-F modification, and (iii) a nucleotide without an internucleotide modification (e.g., a PS modification). In one embodiment, the nucleotide having a 2'OMe modification, the nucleotide having a 2'F modification, and the nucleotide without an internucleotide modification (e.g., a PS modification) are present at any one of nucleotide positions in the [VL domain], for example, (i) the nucleotide position corresponds to any one of 44 to 48 of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to any one of V1 to V27 and 46 to 48 according to CtNS.

[0618] Design Guidance 5 In one embodiment, the TREM comprises a sequence of Formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], where, independently, [L1] and [VL domain] are optional, and one of the [TH domains] comprises a nucleotide with a non-naturally occurring modification. In one embodiment, the nucleotide with the non-naturally occurring modification is present at any one of nucleotide positions 49-65, e.g., (i) nucleotide positions corresponding to 49-65 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 49-65 according to CtNS. In one embodiment, the [TH domain] comprises a nucleotide with a 2'-nucleotide sugar modification. In one embodiment, the nucleotide having a 2'-nucleotide sugar modification is at any one of nucleotide positions 49-65, for example, (i) nucleotide positions corresponding to 49-65 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 49-65 according to CtNS.

[0619] (a) In one embodiment, the nucleotide having the non-natural modification is at any one of nucleotide positions 49-53 and 61-65, for example, (i) nucleotide positions corresponding to 49-53 and 61-65 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 49-53 and 61-65 according to CtNS.

[0620] (b) In one embodiment, the [TH domain] comprises a nucleotide having a 2'-OMe modification. In one embodiment, the nucleotide having a 2'-OMe modification is at any one of nucleotide positions 49-53, 61-62, and 64-65, e.g., (i) nucleotide positions corresponding to 49-53, 61-62, and 64-65 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 49-53, 61-62, and 64-65 of CtNS.

[0621] (c) In one embodiment, the [TH domain] comprises a nucleotide having a 2'-F modification. In one embodiment, the nucleotide having a 2'-F modification is at nucleotide position 63, e.g., (i) the nucleotide position corresponds to 63 in SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 63 according to CtNS.

[0622] (d) In one embodiment, the [TH domain] comprises a nucleotide that does not have an internucleotide modification (e.g., a PS modification). In one embodiment, the nucleotide that does not have an internucleotide modification (e.g., a PS modification) is at any one of nucleotide positions 49-53 and 61-65, for example, (i) nucleotide positions corresponding to 49-53 and 61-65 of SEQ ID NO: 622, or (ii) nucleotide positions corresponding to 49-53 and 61-65 of CtNS.

[0623] Design Guidance 6 In one embodiment, the TREM comprises a sequence of formula (I): [L1]-[ASt domain 1]-[L2]-[DH domain]-[L3]-[ACH domain]-[VL domain]-[TH domain]-[L4]-[ASt domain 2], where, independently, [L1] and [VL domain] are optional, and each of [DH domain] and [TH domain] comprises a nucleotide having a non-natural modification.

[0624] (a) In one embodiment, the [DH domain] comprises a nucleotide having a non-natural modification at any one of nucleotide positions 14-21 (e.g., 14-20, e.g., 16-18), e.g., (i) nucleotide positions correspond to 14-21 (e.g., 14-20, e.g., 16-18) of SEQ ID NO: 622, or (ii) nucleotide positions correspond to 14-21 (e.g., 14-20, e.g., 16-18) of SEQ ID NO: 622. In one embodiment, the non-natural modification is a 2'-nucleotide sugar modification (e.g., 2'-OMe).

[0625] (b) In one embodiment, the [TH domain] comprises a nucleotide having an unnatural modification at any one of nucleotide positions 54-60, e.g., (i) the nucleotide positions correspond to 54-60 of SEQ ID NO: 622, or (ii) the nucleotide positions correspond to 54-60 by CtNS. In one embodiment, the unnatural modification is a 2'-nucleotide sugar modification (e.g., 2'-F).

[0626] (c) In one embodiment, the [TH domain] comprises a nucleotide having a 2'OMe at any one of nucleotide positions 54-60 (e.g., 54, 56, 57, or 59), e.g., (i) the nucleotide position corresponds to 54-60 (e.g., 54, 56, 57, or 59) of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 54-60 (e.g., 54, 56, 57, or 59) according to CtNS.

[0627] (d) In one embodiment, the [TH domain] comprises a nucleotide having a 2'-F at any one of nucleotide positions 54-60 (e.g., 57, 58, or 60), e.g., (i) the nucleotide position corresponds to 54-60 (e.g., 57, 58, or 60) of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 54-60 (e.g., 57, 58, or 60) according to CtNS.

[0628] (e) In one embodiment, the TREM comprises an unnatural modification in both the [DH domain] and the [TH domain], wherein (i) the [DH domain] comprises a nucleotide having an unnatural modification at any one of nucleotide positions 14-21 (e.g., 14-20, e.g., 16-18), e.g., (i) the nucleotide position corresponds to 14-21 (e.g., 14-20, e.g., 16-18) of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 14-21 (e.g., 14-20, e.g., 16-18) by CtNS, and (ii) the [TH domain] comprises a nucleotide having an unnatural modification at any one of nucleotide positions 54-60, e.g., (i) the nucleotide position corresponds to 54-60 of SEQ ID NO: 622, or (ii) the nucleotide position corresponds to 54-60 by CtNS. In some embodiments, the unnatural modification is selected from a 2'OMe or a 2'F modification.

[0629] Table 21 below summarizes a representative set of TREMs described herein that correlate to the design guidance outlined above, eg, design guidance 1-6. [Table 21-1] [Table 21-2] [Table 21-3]

[0630] TREM, TREM core fragments, and TREM fragment fusions In one embodiment, the TREM, TREM core fragment, or TREM fragment disclosed herein comprises an additional moiety, such as a fusion moiety. In one embodiment, the fusion moiety can be used for purification, to alter the folding of the TREM, TREM core fragment, or TREM fragment, or as a targeting moiety. In one embodiment, the fusion moiety can comprise a tag, a linker, be cleavable, or contain an enzyme binding site. In one embodiment, the fusion moiety can be positioned at the N-terminus of the TREM or the C-terminus of the TREM, TREM core fragment, or TREM fragment. In one embodiment, the fusion moiety can be encoded by the same or a different nucleic acid molecule that encodes the TREM, TREM core fragment, or TREM fragment.

[0631] TREM consensus sequence In one embodiment, the TREM disclosed herein comprises a consensus sequence provided herein.

[0632] In one embodiment, the TREM disclosed herein has formula I ZZZ wherein: ZZZ represents any of the 20 amino acids, and Formula I corresponds to all species.

[0633] In one embodiment, the TREM disclosed herein has formula II ZZZ wherein: ZZZ represents any of the 20 amino acids, and Formula II corresponds to mammals.

[0634] In one embodiment, the TREM disclosed herein has formula III ZZZ wherein: ZZZ represents any of the 20 amino acids, and Formula III corresponds to the human form.

[0635] In one embodiment, ZZZrepresents 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.

[0636] In one embodiment, the TREM disclosed herein comprises a property selected from the following: a) under physiological conditions, residue R0 forms a linker region, e.g., a 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., an AStD stem region, or 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, or e) Under physiological conditions, residue -R 29 form a linker region, e.g., a linker 3 region, or 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, or g) Under physiological conditions, residues -[R 47 ] x comprises, for example, a variable region described herein, or 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 form a linker region, for example, the linker 4 region.

[0637] Alanine TREM consensus sequence In one embodiment, the TREM disclosed herein has formula I ALA (SEQ ID NO: 562), i.e. 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, the consensus for Ala is R = none; R 14 ,R 57 = A or none independently; R 26 = A, C, G or None; R5, R6, R 15 ,R 16 ,R 21 ,R 30 ,R 31 ,R32 ,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 = N or None independently; R 11 ,R 35 ,R 65 = A, C, U or none independently; R1, R9, R 20 ,R 38 ,R 40 ,R 51 ,R 52 ,R 56 = A, G or none independently; R7, R 22 ,R 25 ,R 27 ,R 29 ,R 46 ,R 53 ,R 72 = independently A, G, U or none; R 24 ,R 69 = independently A, U or none; R 70 ,R 71 = independently C or none; R3, R4 = independently C, G or none; R 12 ,R 33 ,R 36 ,R 62 ,R 68 = independently C, G, U or none; R 13 ,R 17 ,R 28 ,R 39 ,R 55 ,R 60 ,R 61 = independently C, U or none; R 10 ,R 19 ,R 23 = G or None independently; R2 = G, U or None; R8, R 18 ,R 54 = independently U or None; [R 47 ]x =N or None, for example, x=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= 70-271,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). However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0638] In one embodiment, the TREM disclosed herein has formula II ALA (SEQ ID NO: 563), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Ala is: R0,R 18 = None; R 14 ,R 24 ,R 57 = independently A or None; R 15 ,R 26,R 64 = independently A, C, G or none; R 16 ,R 31 ,R 50 ,R 59 = N or None independently; R 11 ,R 32 ,R 37 ,R 41 ,R 43 ,R 45 ,R 49 ,R 65 ,R 66 = independently A, C, U or none; R1, R5, R9, R 25 ,R 27 ,R 38 ,R 40 ,R 46 ,R 51 ,R 56 = independently A, G or none; R7,R 22 ,R 29 ,R 42 ,R 44 ,R 53 ,R 63 ,R 72 = independently A, G, U or none; R6,R 35 ,R 69 = independently A, U or None; R 55 ,R 60 ,R 70 ,R 71 = independently C or none; R3 = C, G or None; R 12 ,R 36 ,R 48 = independently C, G, U or none; 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 none; R4,R 10 ,R 19 ,R 20 ,R 23 ,R 52 = independently G or None; R2, R8, R 33 = independently G, U or none; R 21 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0639] In one embodiment, the TREM disclosed herein has formula III ALA (SEQ ID NO: 564), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Ala is: R0,R 18 = None; R 14 ,R 24 ,R 57 ,R 72 = independently A or None; R 15 ,R 26 ,R 64 = independently A, C, G or none; R 16 ,R 31 ,R 50 = N or None independently; R 11 ,R 32 ,R 37 ,R 41 ,R 43 ,R 45 ,R 49 ,R 65 ,R 66 = independently A, C, U or none; R5, R9, R 25 ,R 27 ,R 38 ,R 40 ,R 46 ,R 51 ,R 56 = independently A, G or none; R7,R 22 ,R 29 ,R 42 ,R 44 ,R 53 ,R 63 = independently A, G, U or none; R6,R 35 = independently A, U or None; R 55 ,R 60 ,R 61 ,R 70 ,R 71 = independently C or none; R 12 ,R 48 ,R 59= independently C, G, U or none; R 13 ,R 17 ,R 28 ,R 30 ,R 34 ,R 39 ,R 58 ,R 62 ,R 67 ,R 68 = independently C, U or none; R1, R2, R3, R4, R 10 ,R 19 ,R 20 ,R 23 ,R 52 = independently G or None; R 33 ,R 36 = independently G, U or none; R8,R 21 ,R 54 ,R 69 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0640] Arginine TREM consensus sequence In one embodiment, the TREM disclosed herein has formula I ARG (SEQ ID NO: 565), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Arg is: R 57 =A or None; R9,R 27 = independently A, C, G or none; R1, R2, R3, R4, R5, R6, R7, R 11,R 12 ,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 = N or None independently; R 13 ,R 17 ,R 41 = independently A, C, U or none; R 19 ,R 20 ,R 24 ,R 40 ,R 56 = independently A, G or none; R 14 ,R 15 ,R 72 = independently A, G, U or none; R 18 =A,U or None; R 38 =C or None; R 35 ,R 43 ,R 61 = independently C, G, U or none; R 28 ,R 55 ,R 59 ,R 60 = independently C, U or none; R0,R 10 ,R 52 = independently G or None; R8,R 39 = independently G, U or none; R 36 ,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0641] In one embodiment, the TREM disclosed herein has formula II ARG(Array number 566), that is 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<N 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<00024LL>-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 It should be noted that in the translation, for the 7-digit tags like 40 , 44 , etc., they are kept exactly as in the original according to the requirements. If there are any inaccuracies in the original text understanding, please let me know.-R 72 contains an array of where R is a ribonucleotide residue and the consensus for Arg is: R 18 = None; R 24 ,R 57 = independently A or None; R 41 =A,C or None; R3, R7, R 34 ,R 50 = independently A, C, G or none; R2, R5, R6, R 12 ,R 26 ,R 32 ,R 37 ,R 44 ,R 58 ,R 66 ,R 67 ,R 68 ,R 70 = N or None independently; R 49 ,R 71 = independently A, C, U or none; R1,R 15 ,R 19 ,R 25 ,R 27 ,R 40 ,R 45 ,R 46 ,R 56 ,R 72 = independently A, G or none; R 14 ,R 29 ,R 63 = independently A, G, U or none; R 16 ,R 21 = independently A, U or None; R 38 ,R 61 = independently C or none; R 33 ,R 48 = independently C, G or none; R4, R9, R 11 ,R 43 ,R 62 ,R 64 ,R 69= independently C, G, U or none; R 13 ,R 22 ,R 28 ,R 30 ,R 31 ,R 35 ,R 55 ,R 60 ,R 65 = independently C, U or none; R0,R 10 ,R 20 ,R 23 ,R 51 ,R 52 = independently G or None; R8,R 39 ,R 42 = independently G, U or none; R 17 ,R 36 ,R 53 ,R 54 ,R 59 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0642] In one embodiment, the TREM disclosed herein has formula III ARG (SEQ ID NO: 567), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Arg is: R 18 = None; R 15 ,R 21 ,R 24 ,R 41 ,R 57 = independently A or None; R 34 ,R44 = independently A, C or none; R3, R5, R 58 = independently A, C, G or none; R2, R6, R 66 ,R 70 = N or None independently; R 37 ,R 49 = independently A, C, U or none; R1,R 25 ,R 29 ,R 40 ,R 45 ,R 46 ,R 50 = independently A, G or none; R 14 ,R 63 ,R 68 = independently A, G, U or none; R 16 =A,U or None; R 38 ,R 61 = independently C or none; R7,R 11 ,R 12 ,R 26 ,R 48 = independently C, G or none; R 64 ,R 67 ,R 69 = independently C, G, U or none; 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 none; R0,R 10 ,R 19 ,R 20 ,R 23 ,R 27 ,R 33 ,R 51 ,R 52 ,R 56 ,R72 = independently G or None; R8, R9, R 32 ,R 39 ,R 42 = independently G, U or none; R 17 ,R 36 ,R 53 ,R 54 ,R 59 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0643] Asparagine TREM consensus sequence In one embodiment, the TREM disclosed herein has formula I ASN (SEQ ID NO: 568), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Asn is: R0,R 18 = None; R 41 =A or None; R 14 ,R 48 ,R 56 = independently A, C, G or none; 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 = N or None independently; R 11 ,R 13 ,R 22 ,R 42 ,R 55 ,R 59 = independently A, C, U or none; R9,R 15 ,R 24 ,R 27 ,R 34 ,R 37 ,R 51 ,R 72 = independently A, G or none; R1, R7, R 25 ,R 69 = independently A, G, U or none; R 40 ,R 57 = independently A, U or None; R60 =C or None; R 33 =C,G or None; R 21 ,R 32 ,R 43 ,R 64 = independently C, G, U or none; R3,R 16 ,R 28 ,R 35 ,R 36 ,R 61 = independently C, U or none; R 10 ,R 19 ,R 20 ,R 52 = independently G or None; R 54 =G,U or None; R8,R 23 ,R 38 ,R 39 ,R 53 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0644] In one embodiment, the TREM disclosed herein has formula II ASN (SEQ ID NO: 569), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Asn is: R0,R 18 = None R 24 ,R 41 ,R 46 ,R 62 = independently A or None; R 59 =A,C or None; R 14 ,R 56 ,R 66 = independently A, C, G or none; R 17 ,R 29 = N or None independently; R 11 ,R 26 ,R 42 ,R 55 = independently A, C, U or none; 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 none; R 44 ,R 45 ,R 58 = independently A, G, U or none; R 40 ,R 57 = independently A, U or None; R5,R 28 ,R 60 = independently C or none; R 33 ,R 65 = independently C, G or none; R 21 ,R 43 ,R 71 = independently C, G, U or none; R3, R6, R 13 ,R 22 ,R 32 ,R 35 ,R 36 ,R 61 ,R 63 ,R 64 = independently C, U or none; R7,R 10 ,R 19 ,R 20 ,R 27 ,R 49 ,R 52= independently G or None; R 54 =G,U or None; R2, R4, R8, R 16 ,R 23 ,R 30 ,R 31 ,R 38 ,R 39 ,R 50 ,R 53 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0645] In one embodiment, the TREM disclosed herein has formula III ASN (SEQ ID NO: 570), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Asn is: R0,R 18 = None R 24 ,R 40 ,R 41 ,R 46 ,R 62 = independently A or None; R 59 =A,C or None; R 14 ,R 56 ,R 66 = independently A, C, G or none; R 11 ,R 26 ,R 42 ,R 55 = independently A, C, U or none; 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 none; R 44 ,R 45 ,R 58 = independently A, G, U or none; R 57 =A,U or None; R5,R 28 ,R 60 = independently C or none; R 33 ,R 65 = independently C, G or none; R 17 ,R 21 ,R 29 = independently C, G, U or none; R3, R6, R 13 ,R 22 ,R 32 ,R 35 ,R36 ,R 43 ,R 61 ,R 63 ,R 64 ,R 71 = independently C, U or none; R7,R 10 ,R 19 ,R 20 ,R 25 ,R 27 ,R 49 ,R 52 ,R 72 = independently G or None; R 54 =G,U or None; R2, R4, R8, R 16 ,R 23 ,R 30 ,R 31 ,R 38 ,R 39 ,R 50 ,R 53 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0646] Aspartic acid TREM consensus sequence In one embodiment, the TREM disclosed herein has formula I ASP (SEQ ID NO: 571), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Asp is: R0=none R 24 ,R 71 = independently A, C or none; R 33 ,R 46= independently A, C, G or none; 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 = N or None independently; R 13 ,R 21 ,R 34 ,R 41 ,R 57 ,R 65 = independently A, C, U or none; 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 none; R7,R 25 ,R 42 = independently A, G, U or none; R 39 =C or None; R 50 ,R 62 = independently C, G or none; R 30 ,R 43 ,R 45 ,R 55 ,R 70 = independently C, G, U or none; R8,R 11 ,R 17 ,R 18 ,R 28 ,R 35 ,R 53 ,R 59 ,R 60 ,R61 = independently C, U or none; R 19 ,R 52 = independently G or None; R1=G,U or None; R 23 ,R 36 ,R 38 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0647] In one embodiment, the TREM disclosed herein has formula II ASP (SEQ ID NO: 572), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Asp is: R0,R 17 ,R 18 ,R 23 = independently none; R9,R 40 = independently A or None; R 24 ,R 71 = independently A, C or none; R 67 ,R 68 = independently A, C, G or none; R2, R6, R 66 = N or None independently; R 57 ,R 63 = independently A, C, U or none; 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 none; R7,R 12 ,R 26 ,R 65 = independently A, U or None; R 39 ,R 61 ,R 62 = independently C or none; R3,R 31 ,R 45 ,R 70 = independently C, G or none; R4, R5, R 29 ,R 43 ,R 55 = independently C, G, U or none; R8,R 11 ,R 13 ,R 30 ,R 32 ,R34 ,R 35 ,R 41 ,R 48 ,R 53 ,R 59 ,R 60 = independently C, U or none; R 15 ,R 19 ,R 20 ,R 25 ,R 42 ,R 50 ,R 52 = independently G or None; R1,R 22 ,R 49 ,R 58 ,R 69 = independently G, U or none; R 16 ,R 21 ,R 28 ,R 36 ,R 38 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0648] In one embodiment, the TREM disclosed herein has formula III ASP (SEQ ID NO: 573), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Asp is: R0,R 17 ,R 18 ,R 23 = None R9,R 12 ,R 40 ,R 65 ,R 71 = independently A or None; R2,R 24 ,R 57 = independently A, C or none; R6,R 14 ,R 27 ,R 46 ,R 51 ,R 56 ,R 64 ,R 67 ,R 68 = independently A, G or none; R3,R 31 ,R 35 ,R 39 ,R 61 ,R 62 = independently C or none; R 66 =C,G or None; 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 none; 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 None; R 22 ,R 58 = independently G, U or none; R1, R4, R7, R 11 ,R 13 ,R 16 ,R 21 ,R 26 ,R 28 ,R 36 ,R 38,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0649] Cysteine ​​TREM consensus sequence In one embodiment, the TREM disclosed herein has formula I CYS (SEQ ID NO: 574), i.e. R0-R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R12 -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 contains an array of where R is a ribonucleotide residue and the consensus for Cys is: R0=none R 14 ,R 39 ,R 57 = independently A or None; R 41 =A,C or None; R 10 ,R 15 ,R 27 ,R 33 ,R 62 = independently A, C, G or none; 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 = N or None independently; R 65 =A,C,U or None; R9,R 25 ,R 37 ,R 40 ,R 52 ,R 56 = independently A, G or none; R7,R 20 ,R 51 = independently A, G, U or none; R 18 ,R 38 ,R 55 = independently C or none; R2 = C, G or none; R 21 ,R 28 ,R 43 ,R 50 = independently C, G, U or none; R 11 ,R 22 ,R 23 ,R 35 ,R 36 ,R 59 ,R 60 ,R 61 ,R 71 ,R 72 = independently C, U or none; R1,R 19 = independently G or None; R 17 =G,U or None; R8,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0650] In one embodiment, the TREM disclosed herein has formula II CYS (SEQ ID NO: 575), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Cys is: R0,R 18 ,R 23 = None; R 14 ,R 24 ,R 26 ,R 29 ,R 39 ,R 41 ,R 45 ,R 57 = independently A or None; R 44 =A,C or None; R 27 ,R 62 = independently A, C, G or none; R 16 =A,C,G,U or None; R 30 ,R 70 = independently A, C, U or none; 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 none; R 20 ,R 51 = independently A, G, U or none; R 35 ,R 38 ,R 43 ,R 55 ,R 69 = independently C or none; R2, R4, R 15 = independently C, G or none; R13 =C,G,U or None; 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 none; R1, R3, R 10 ,R 19 ,R 33 ,R 63 = independently G or None; R8,R 17 ,R 21 ,R 64 = independently G, U or none; R 12 ,R 22 ,R 31 ,R 32 ,R 42 ,R 53 ,R 54 ,R 65 = independently U or None; R 59 =U, or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0651] In one embodiment, the TREM disclosed herein has formula III CYS (SEQ ID NO: 576), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Cys is: R0,R 18 ,R 23 = None R 14 ,R 24 ,R 26 ,R 29 ,R 34 ,R 39 ,R41 ,R 45 ,R 57 ,R 58 = independently A or None; R 44 ,R 70 = independently A, C or none; R 62 =A,C,G or None; R 16 =N or None; R5, R7, R9, R 20 ,R 40 ,R 46 ,R 51 ,R 52 ,R 56 ,R 66 = independently A, G or none; R 28 ,R 35 ,R 38 ,R 43 ,R 55 ,R 67 ,R 69 = independently C or none; R4,R 15 = independently C, G or none; R6,R 11 ,R 13 ,R 30 ,R 48 ,R 49 ,R 50 ,R 60 ,R 61 ,R 68 ,R 71 ,R 72 = independently C, U or none; R1, R2, R3, R 10 ,R 19 ,R 25 ,R 27 ,R 33 ,R 37 ,R 63 = independently G or None; R8,R 21 ,R 64 = independently G, U or none; R 12 ,R 17 ,R 22 ,R 31 ,R 32 ,R36 ,R 42 ,R 53 ,R 54 ,R 59 ,R 65 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0652] Glutamine TREM consensus sequence In one embodiment, the TREM disclosed herein has formula I GLN (SEQ ID NO: 577), i.e. 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 including the arrangement of​ where R is a ribonucleotide residue and the consensus for Gln is: R0,R 18 = None; R 14 ,R 24 ,R 57 = independently A or None; R9,R 26 ,R 27 ,R 33 ,R 56 = independently A, C, G or none; 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 = N or None independently; R 17 ,R 23 ,R 43 ,R 65 ,R 71 = independently A, C, U or none; R 15 ,R 40 ,R 51 ,R 52 = independently A, G or none; R1, R7, R 72 = independently A, G, U or none; R3,R 11 ,R 37 ,R 60 ,R 64= independently C, G, U or none; R 28 ,R 35 ,R 55 ,R 59 ,R 61 = independently C, U or none; R 10 ,R 19 ,R 20 = independently G or None; R 39 =G,U or None; R8,R 36 ,R 38 ,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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=70-271 ,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) However, TREMs have one or both of the following properties: less than 15% of the residues are N, or no more than 20 residues are present.

[0653] In one embodiment, the TREM disclosed herein has formula II GLN (SEQ ID NO: 578), i.e. 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 contains an array of where R is a ribonucleotide residue and the consensus for Gln is: R0,R 18 ,R 23 = None R 14 ,R 24 ,R 57 = independently A or None; R 17 ,R 71 = independently A, C or none; R 25 ,R 26 ,R 33 ,R 44 ,R 46 ,R 56 ,R 69 = independently A, C, G or none; R4, R5, R 12 ,R 22 ,R 29 ,R 30 ,R 48 ,R 49 ,R 63 ,R 67 ,R 68 = N or None independently; R 31 ,R 43 ,R 62 ,R 65 ,R 70 = independently A, C, U or none; R 15 ,R 27 ,R 34 ,R 40 ,R 41 ,R 51 ,R 52 = independently A, G or none; R2, R7, R 21 ,R 45 ,R 50 ,R 58 ,R66 ,R 72 = independently A, G, U or none; R3,R 13 ,R 32 ,R 37 ,R 42 ,R 60 ,R 64 = independently C, G, U or none; R6,R 11 ,R 28 ,R 35 ,R 55 ,R 59 ,R 61 = independently C, U or none; R9,R 10 ,R 19 ,R 20 = independently G or None; R1,R 16 ,R 39 = independently G, U or none; R8,R 36 ,R 38 ,R 53 ,R 54 = independently U or None; [R 47 ] x =N or None; 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...

Claims

1. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1] -[ASt domain 1] -[L2] -[DH domain] -[L3] -[ACH domain] -[VL domain] -[TH domain] -[L4] -[ASt domain 2] (I), During the ceremony, independently, [L1] and [VL domain] are optional; The TREM, wherein the nucleotide within [L1]-[AST domain 1]-[L2] includes a nucleotide having a non-natural modification.

2. 10. The TREM of claim 1, wherein the non-natural modification is present on the 2' position of the nucleotide sugar or within an internucleotide region.

3. 2. The TREM of claim 1, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), a 2'-halo (2'F or 2'Cl), a 2'-O-methoxyethyl (2'MOE), a 2'-deoxy modification, or a phosphorothioate modification.

4. 2. The TREM of claim 1, wherein the TREM is a TREM shown in FIG. 2 or has a sequence selected from the sequences shown in FIG.

5. The TREM is (a) a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2; (b) a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2; and / or (c) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications compared to the TREM shown in Figure 2 2. The TREM of claim 1, comprising:

6. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of nucleotides 1-9 of SEQ ID NO: 622; (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622; and / or (iii) the TREM comprises the nucleotide sequence of SEQ ID NO: 622; 10. The TREM of claim 1.

7. The TREM of claim 1, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623 or 624.

8. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1] -[ASt domain 1] -[L2] -[DH domain] -[L3] -[ACH domain] -[VL domain] -[TH domain] -[L4] -[ASt domain 2] (I), During the ceremony, independently, [L1] and [VL domain] are optional; The TREM, wherein the nucleotide in [DH domain 1] or [L3] comprises a nucleotide having a non-natural modification.

9. 9. The TREM of claim 8, wherein the non-natural modification is on the 2' position of the nucleotide sugar or within an internucleotide region.

10. 9. The TREM of claim 8, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), a 2'-halo (2'F or 2'Cl), a 2'-O-methoxyethyl (2'MOE), a 2'-deoxy modification, or a phosphorothioate modification.

11. 9. The TREM of claim 8, wherein the TREM is a TREM shown in FIG. 2 or has a sequence selected from the sequences shown in FIG.

12. The TREM is (a) a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2; (b) a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2; and / or (c) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications compared to the TREM shown in Figure 2 9. The TREM of claim 8, comprising:

13. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of nucleotides 10-26 of SEQ ID NO: 622; (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622; and / or (iii) the TREM comprises the nucleotide sequence of SEQ ID NO: 622; 9. The TREM of claim 8.

14. The TREM of claim 8, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623 or 624.

15. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1] -[ASt domain 1] -[L2] -[DH domain] -[L3] -[ACH domain] -[VL domain] -[TH domain] -[L4] -[ASt domain 2] (I), During the ceremony, independently, [L1] and [VL domain] are optional; The TREM, wherein the nucleotides in the ACH domain include nucleotides having a non-natural modification.

16. 16. The TREM of claim 15, wherein the non-natural modification is on the 2' position of the nucleotide sugar or within an internucleotide region.

17. 16. The TREM of claim 15, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), a 2'-halo (2'F or 2'Cl), a 2'-O-methoxyethyl (2'MOE), a 2'-deoxy modification, or a phosphorothioate modification.

18. 16. The TREM of claim 15, wherein the TREM is a TREM shown in Figure 2 or has a sequence selected from the sequences shown in Figure 2.

19. The TREM is (a) a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2; (b) a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2; and / or (c) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications compared to the TREM shown in Figure 2 16. The TREM of claim 15, comprising:

20. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of nucleotides 27-43 of SEQ ID NO: 622; (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622; and / or (iii) the TREM comprises the nucleotide sequence of SEQ ID NO: 622; 16. The TREM of claim 15.

21. The TREM of claim 15, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623 or 624.

22. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1] -[ASt domain 1] -[L2] -[DH domain] -[L3] -[ACH domain] -[VL domain] -[TH domain] -[L4] -[ASt domain 2] (I), During the ceremony, independently, [L1] and [VL domain] are optional; The TREM, wherein the nucleotide in the VL domain comprises a nucleotide having a non-natural modification.

23. 23. The TREM of claim 22, wherein the non-natural modification is on the 2' position of the nucleotide sugar or within an internucleotide region.

24. 23. The TREM of claim 22, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), a 2'-halo (2'F or 2'Cl), a 2'-O-methoxyethyl (2'MOE), a 2'-deoxy modification, or a phosphorothioate modification.

25. 23. The TREM of claim 22, wherein the TREM is a TREM shown in Figure 2 or has a sequence selected from the sequences shown in Figure 2.

26. The TREM is (a) a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2; (b) a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2; and / or (c) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications compared to the TREM shown in Figure 2 23. The TREM of claim 22, comprising:

27. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of nucleotides 44-48 of SEQ ID NO: 622; (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622; and / or (iii) the TREM comprises the nucleotide sequence of SEQ ID NO: 622; 23. The TREM of claim 22.

28. 23. The TREM of claim 22, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623 or 624.

29. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1] -[ASt domain 1] -[L2] -[DH domain] -[L3] -[ACH domain] -[VL domain] -[TH domain] -[L4] -[ASt domain 2] (I), During the ceremony, independently, [L1] and [VL domain] are optional; The TREM, wherein the nucleotide in the TH domain comprises a nucleotide having a non-natural modification.

30. 30. The TREM of claim 29, wherein the non-natural modification is on the 2' position of the nucleotide sugar or within an internucleotide region.

31. 30. The TREM of claim 29, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), a 2'-halo (2'F or 2'Cl), a 2'-O-methoxyethyl (2'MOE), a 2'-deoxy modification, or a phosphorothioate modification.

32. 30. The TREM of claim 29, wherein the TREM is a TREM shown in Figure 2 or has a sequence selected from the sequences shown in Figure 2.

33. The TREM is (a) a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2; (b) a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2; and / or (c) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications compared to the TREM shown in Figure 2 30. The TREM of claim 29, comprising:

34. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of nucleotides 49-65 of SEQ ID NO: 622; (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622; and / or (iii) the TREM comprises the nucleotide sequence of SEQ ID NO: 622; 30. The TREM of claim 29.

35. 30. The TREM of claim 29, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623 or 624.

36. A tRNA effector molecule (TREM) comprising a sequence of formula (I): [L1] -[ASt domain 1] -[L2] -[DH domain] -[L3] -[ACH domain] -[VL domain] -[TH domain] -[L4] -[ASt domain 2] (I), During the ceremony, independently, [L1] and [VL domain] are optional; The TREM, wherein the nucleotide in [L4] or [AST domain 2] comprises a nucleotide having a non-natural modification.

37. 37. The TREM of claim 36, wherein the non-natural modification is on the 2' position of the nucleotide sugar or within an internucleotide region.

38. 37. The TREM of claim 36, wherein the non-natural modification is selected from a 2'-O-methyl (2-OMe), a 2'-halo (2'F or 2'Cl), a 2'-O-methoxyethyl (2'MOE), a 2'-deoxy modification, or a phosphorothioate modification.

39. 37. The TREM of claim 36, wherein the TREM is a TREM shown in Figure 2 or has a sequence selected from the sequences shown in Figure 2.

40. The TREM is (a) a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity to a TREM shown in Figure 2; (b) a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the TREM shown in Figure 2; and / or (c) 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-natural modifications compared to the TREM shown in Figure 2 37. The TREM of claim 36, comprising:

41. (i) the non-natural modification is present at a nucleotide position corresponding to one or more of nucleotides 66-76 of SEQ ID NO: 622; (ii) the TREM differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides compared to the nucleotide sequence of SEQ ID NO: 622; and / or (iii) the TREM comprises the nucleotide sequence of SEQ ID NO: 622; 37. The TREM of claim 36.

42. 37. The TREM of claim 36, wherein the TREM comprises the nucleotide sequence of SEQ ID NO: 623 or 624.

43. TREM according to design guidance 1, 2, 3, 4, 5, or 6 described herein.

44. TREM according to any two, three, four, five, or all of design guidance 1, 2, 3, 4, 5, and 6 described herein.

45. A pharmaceutical composition comprising the TREM described in claim 1.

46. 46. ​​A lipid nanoparticle formulation comprising the TREM of claim 1 or the pharmaceutical composition of claim 45.

47. 10. A composition for use in a method of treating a subject having a disease or disorder associated with PTC, the composition comprising the TREM of claim 1, or a core fragment or fragment thereof.

48. 48. The composition of claim 47, wherein the PTC-associated disease or disorder comprises hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease.