Non-LTR retrotransposon systems and their use

Abstract

Non-LTR retrotransposon systems are provided. Isolated functional proteins encoded by retrotransposons, nucleic acids encoding functional proteins, nucleic acid sets, nucleic acid constructs, compositions, recombinant vectors, recombinant host cells, and kits are provided. Methods for introducing exogenous nucleic acid fragments into the genome of a host cell, methods for editing the genome of a host cell, and methods for obtaining a host cell containing exogenous nucleic acid fragments in its genome are also provided. Further, the use of functional proteins encoded by retrotransposons, nucleic acids, nucleic acid sets, nucleic acid constructs, compositions, recombinant vectors, or recombinant host cells for introducing exogenous nucleic acid fragment genes into the genome of a host cell, or for preparing drugs or formulations, is provided.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to Chinese Patent Application No. 202310380978.4, filed with the State Intellectual Property Administration of China on 11 April 2023, the entire contents of which are incorporated herein by reference for all purposes.

[0002] This application relates to the field of molecular biology, and more specifically to non-LTR retrotransposon systems and their use. More specifically to isolated functional proteins encoded by retrotransposons, nucleic acids encoding functional proteins, nucleic acid sets and nucleic acid constructs, and compositions, recombinant vectors, recombinant host cells and kits comprising functional proteins. More specifically to methods for introducing exogenous nucleic acid fragments into the genome of host cells, methods for editing the genome of host cells, and methods for obtaining host cells containing exogenous nucleic acid fragments in their genome. More specifically to the use of functional proteins, nucleic acids, nucleic acid sets, nucleic acid constructs, compositions, recombinant vectors or recombinant host cells for introducing exogenous nucleic acid fragment genes into the genome of host cells, or for preparing drugs or formulations for gene therapy, cell therapy, genome research, or stem cell induction and post-induction differentiation. [Background technology]

[0003] Transposons are DNA sequences (Mobile DNA III) that can be inserted into or removed from a genome to replace their own sequence or a complete copy of their sequence within or between genomes. Transposons are mainly divided into two categories. Of these, type I retrotransposons use RNA as an intermediate. In the process of moving from a donor site to a new insertion site, type I retrotransposons require host RNA polymerase to transcribe the transposable element into RNA, and then the reverse transcriptase domain encoded by the retrotransposon reverse transcribes the RNA copy of the transposable element into the DNA to be inserted into the new site. During the retrotransposition process of this type of retrotransposon, the sequence at the donor site is not cleaved from its original site, so multiple copies of the retrotransposable element may exist in the genome.

[0004] Based on their different replication and integration mechanisms, retrotransposons can be divided into two categories: LTR (long terminal repeat) and non-LTR. The latter are a type of mobile genetic element widely distributed in eukaryotic cell genomes. Based on the characteristics of the nuclease functional domains they contain, non-LTR retrotransposons can be further divided into two categories: aprine / apyrimidine nucleases (APE type) and restriction enzyme-like nucleases (RLE type). Among these, the R2 family of retrotransposons are the main representatives of RLE-type non-LTR retrotransposons. Some R2 retrotransposons exhibit high target site specificity and are specifically integrated into genes encoding 28s rRNA.

[0005] Gene insertion and large fragment integration have significant application value in fields such as gene therapy, molecular breeding of plants and animals, and the manipulation of industrial microorganisms. Currently, the industry lacks effective tools and systems for the insertion and integration of large fragment genes. In recent years, the scientific community has developed several tools and methods that can insert and integrate large fragment genes, but these methods still have several problems. For example, in cell immunotherapy and gene therapy for genetic diseases, lentiviruses or retroviruses are most commonly used to incorporate gene sequences, and based on this, there are several therapeutic products for the treatment of tumors and genetic disorders (Aiuti, A., Roncarolo, MG and Naldini, L. (2017) Gene therapy for ADA-SCID, the first marketing approval of an ex vivo gene therapy in Europe: paving the road for the next generation of advanced therapy medicinal products. EMBO Mol. Med. 9, 737-740; Aiuti, A. et al. (2009) Gene therapy for immunodeficiency due to adenosine deaminase deficiency. N.Engl. J. Med. 360, 447-458). However, using viruses to incorporate large fragment genes has several potential limitations in terms of application: firstly, the randomness of viral integration in the genome poses a risk of cancer; secondly, the size of exogenous genes that a virus can possess is also limited, making it unsuitable for the transfer of large fragment genes for therapeutic purposes; thirdly, the immunogenicity of the virus may affect the long-term expression and re-administration of exogenous therapeutic genes; and fourthly, the production of viruses requires the assistance of living cells, which makes quality control and downstream processing of such products more complex and expensive, thus having certain drawbacks from an industrialization standpoint.Therefore, the integration of large non-viral fragments can avoid various drawbacks caused by viral integration and can become a valuable tool in gene therapy.

[0006] As a non-viral gene integration tool, retrotransposons can achieve integration into the host genome via RNA delivery and stable expression of large fragments of exogenous genes, which can not only reduce adverse effects such as immunogenicity, but also directly treat target cells in vivo in combination with LNPs and other delivery technologies. Due to the lack of relevant retrotransposon tools, there is little application of retrotransposon technology in gene therapy or other fields. Since there are few companies in the world that are mining a large number of active retrotransposons and developing them into therapeutic tools, it is necessary to quickly discover and obtain more active retrotransposon products, verify and detect their functions, and provide more options for the development of gene therapy strategies.

[0007] Note that the methods described in this section are not necessarily methods that have been previously conceived or adopted. Unless otherwise specified, none of the methods described in this section should be regarded as prior art solely because they are included in this section. Similarly, the problems mentioned in this section should not be regarded as being generally recognized in any prior art unless specifically indicated.

Summary of the Invention

Means for Solving the Problems

[0008] Based on this, in order to seek more advanced and effective non-viral gene integration tools, this application provides a non-LTR retrotransposon system and its use. More specifically, this application provides an isolated functional protein encoded by a retrotransposon, having a functional protein sequence selected from (i) below or a variant sequence of the aforementioned functional protein having the functional protein activity of (ii) to (iv): (i) at least one amino acid sequence shown in any one of SEQ ID NOs: 1 to 94; (ii) at least one sequence having deletion, substitution, insertion, or mutation of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids of the amino acid sequence shown in any one of SEQ ID NOs: 1 to 94; (iii) at least one amino acid sequence having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence shown in any one of SEQ ID NOs: 1 to 94; and (iv) at least one sequence obtained by further fusing the amino acid sequence shown in any one of SEQ ID NOs: 1 to 94 with another sequence. The functional proteins encoded by the retrotransposons provided in this application possess high retrotransposition activity and can offer more options for the development of gene integration tools.

[0009] According to one embodiment of this application, an isolated functional protein encoded by a retrotransposon, wherein the functional protein is of formula (I): C (X1) a C (X2) b H (X3) c H (I), A functional protein can be provided that contains the amino acid sequence shown in (of which a, b, and c are the number of amino acids, C is cysteine, H is histidine, (X1) is any amino acid, a is 1, 2, 3, or 4, (X2) is any amino acid, b is 11, 12, 13, 14, 15, 16, or 17, (X3) is any amino acid, and c is 4).

[0010] According to one embodiment of the present application, an isolated functional protein encoded by a retrotransposon, wherein the functional protein has the formula (II): G (X4) d Q G D (X5) e S (X6) f F (X7) g D (II), (wherein d, e, f, and g are the number of amino acids, D is aspartic acid, F is phenylalanine, G is glycine, Q is glutamine, S is serine, (X4) is any amino acid, d is 2, (X5) is any amino acid, e is 2, (X6) is any amino acid, f is 3, (X7) is any amino acid, and g is 30, 31, 32, 33, 34, 35, or 36), a functional protein comprising the amino acid sequence shown can be provided.

[0011] According to one embodiment of the present application, an isolated functional protein, wherein the functional protein has the formula (III): C (X8) h C (X9) i E (X 10 ) j H (X 11 ) k C (X 12 ) l R H (X 13 ) m P D (X 14 ) n (X 15 ) (X 16 ) o K (X 17 ) p Y (III), (Of these, h, i, j, k, l, m, n, o and p are the number of amino acids, C is cysteine, D is aspartic acid, E is glutamic acid, H is histidine, K is lysine, R is arginine, P is proline, Y is tyrosine, (X8) is any amino acid, h is 2, 3 or 4, (X9) is any amino acid, i is 3, 4, 5, 6, 7, 8, 9 or 10, (X 10 ) is any amino acid, j is 3, and (X 11 ) is any amino acid, k is 4, and (X 12 ) is any amino acid, l is 9, and (X 13 ) is any amino acid, m is 31, 32, 33, 34, 35 or 36, (X 14 ) is any amino acid, where n is 11, 12, 13, 14, 15, 16, 17, 18 or 19, (X 15 ) is aspartic acid or glutamic acid, (X 16 ) is any amino acid, o is 15, 16, 17, 18 or 19, (X 17 A functional protein can be provided that contains the amino acid sequence shown in (where p is any amino acid and p is 3).

[0012] According to one embodiment of this application, a functional protein is provided which is an isolated functional protein encoded by a retrotransposon and comprises at least two of the amino acid sequences shown in formulas (I), (II), and (III).

[0013] According to one embodiment of this application, an isolated functional protein encoded by a retrotransposon is provided, comprising the amino acid sequences shown in formulas (I), (II), and (III).

[0014] According to one embodiment of this application, nucleic acids encoding the functional protein described in this application can be provided.

[0015] According to one embodiment of this application, a nucleic acid set can be provided, the nucleic acid set comprising a 5' untranslated region, the 5' untranslated region comprising at least one of the nucleotide sequences shown in SEQ ID NOs. 95 to 188.

[0016] According to one embodiment of the present application, a nucleic acid set can be provided, the nucleic acid set comprising a 3' untranslated region, the 3' untranslated region comprising at least one of the nucleotide sequences shown in SEQ ID NOs: 189-282.

[0017] According to one embodiment of the present application, a nucleic acid set can be provided, the nucleic acid set comprising a 5' untranslated region and a 3' untranslated region, the 5' untranslated region comprising a nucleotide sequence or a variant thereof shown in any one of SEQ ID NOs. 95 to 188, and the 3' untranslated region comprising a nucleotide sequence or a variant thereof shown in any one of SEQ ID NOs. 189 to 282, wherein RNA transcribed from the nucleic acid set can bind to a functional protein encoded by a specific retrotransposon.

[0018] According to one embodiment of this application, a nucleic acid construct can be provided, which comprises the nucleic acid and / or nucleic acid set described in this application.

[0019] According to one embodiment of the present application, a composition may be provided which comprises a functional protein or functional fragment thereof encoded by an R2 family retrotransposon, or a nucleic acid encoding a functional protein or functional fragment thereof, wherein the functional protein or functional fragment thereof has the function of catalyzing the insertion of an exogenous nucleic acid fragment into the genome of a cell; and a nucleic acid set wherein the nucleic acid set can be recognized by a functional protein or functional fragment thereof encoded by a particular retrotransposon.

[0020] According to one embodiment of this application, a recombinant vector can be provided, which comprises a nucleic acid encoding a functional protein described in this application, a nucleic acid and nucleic acid set described in this application, a nucleic acid construct described in this application, or a composition described in this application.

[0021] According to one embodiment of this application, recombinant host cells can be provided, which comprise a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid and nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, or a recombinant vector described in this application.

[0022] According to one embodiment of this application, a method can be provided for introducing an exogenous nucleic acid fragment into the genome of a host cell, the method comprising delivering a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid and nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, or a recombinant vector described in this application to a host cell.

[0023] According to one embodiment of the present application, a method for editing the genome of a host cell can be provided, the method comprising delivering a functional protein described in the present application, a nucleic acid encoding a functional protein described in the present application, a nucleic acid and nucleic acid set described in the present application, a nucleic acid construct described in the present application, a composition described in the present application, or a recombinant vector described in the present application to a host cell.

[0024] According to one embodiment of the present application, a method can be provided for obtaining a host cell containing an exogenous nucleic acid fragment in its genome, the method comprising delivering a functional protein described in the present application, a nucleic acid encoding a functional protein described in the present application, a nucleic acid described in the present application, a nucleic acid set described in the present application, a nucleic acid construct described in the present application, a composition described in the present application, or a recombinant vector described in the present application to a host cell.

[0025] According to one embodiment of this application, it is possible to provide the use of a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid described in this application, a nucleic acid construct described in this application, a composition described in this application, a recombinant vector described in this application, or a recombinant host cell described in this application for introducing an exogenous nucleic acid fragment into the genome of a host cell.

[0026] According to one embodiment of this application, it is possible to provide the use of the functional protein described in this application, the nucleic acid encoding the functional protein described in this application, the nucleic acid described in this application, the nucleic acid set described in this application, the nucleic acid construct described in this application, the composition described in this application, the recombinant vector described in this application, or the recombinant host cell described in this application for gene therapy, cell therapy, genome research, or induction and post-induction differentiation of stem cells.

[0027] According to one embodiment of this application, a kit can be provided which comprises a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid described in this application, a nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, a recombinant vector described in this application, or a recombinant host cell described in this application.

[0028] It should be understood that the information in this section is not intended to identify any important or significant features of the example of this application, nor is it intended to limit the scope of this application. Other features of this application will be readily apparent from the following description.

[0029] The accompanying drawings illustrate embodiments and form part of this specification and are used together with the description herein to illustrate exemplary implementations of the embodiments. The embodiments shown are for illustrative purposes only and do not limit the scope of the claims. Throughout the accompanying drawings, the same reference numerals indicate similar elements, but are not necessarily identical. [Brief explanation of the drawing]

[0030] [Figure 1] Figure 1 shows a schematic diagram of the order of elements in the vector in Example 1. [Figure 2] Figure 2 shows the results of detecting retrotranspositional activity by amplifying the 3' junction of the 28s rRNA gene with a retrotransposon. Different lanes in Figure 2 correspond to different retrotransposons in Example 2 (sample codes 1-100 in Table 1). [Figure 3] Figure 3 shows the detection results of retrotransposition activity using a single copy gene (36b4) as a reference. Different lanes in Figure 3 correspond to different retrotransposons in Example 3 (sample codes 3, 4, 5, 7, 11, 14, 19, 20, 21, 23, 26, 28, 29, 43, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 59, 60, 63, 75, 83, 87, and 98 in Table 1). [Figure 4] Figure 4 shows the results of the quantitative analysis of the retrotransposition activity detected in Figure 3. [Figure 5] Figure 5 shows the results of detecting the integrity of retrotransposons by amplifying the 5' junction of 28s rRNA genes located at different loci within the retrotransposon. Different lanes in Figure 5 correspond to different retrotransposons from Example 4 (sample codes 3, 4, 5, 7, 11, 14, 19, 20, 21, 22, 23, 26, 28, 29, 43, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 59, 60, 63, 75, 83, 87, and 98 in Table 1). [Modes for carrying out the invention]

[0031] Unless otherwise indicated or inconsistent with the context, terms or expressions used herein should be read in conjunction with the entirety of this disclosure and to be understood by those skilled in the art. All technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art unless otherwise defined.

[0032] In this application, the terms “nucleic acid” and “polynucleotide” are used interchangeably and refer to polymeric forms of nucleotides of any length, including deoxyribonucleotides, ribonucleotides, combinations thereof, and their analogues.

[0033] In this application, the terms “polypeptide” and “peptide” are used interchangeably and refer to polymers of amino acids of any length. Therefore, polypeptides, oligopeptides, proteins, antibodies, and enzymes are all included in the definition of polypeptide.

[0034] As described in this application, a “fragment” of a sequence refers to a portion of a sequence. For example, a fragment of a nucleic acid sequence refers to a portion of a nucleic acid sequence, and a fragment of an amino acid sequence refers to a portion of an amino acid sequence.

[0035] As described in this application, a “variant” of a sequence is a polynucleotide or polypeptide that is different from the reference polynucleotide or polypeptide but retains essential properties. A typical variant of a polynucleotide has a different nucleic acid sequence from another reference polynucleotide, and this difference in nucleic acid sequence may or may not alter the amino acid sequence of the polypeptide encoded by the reference polynucleotide. A typical variant of a polypeptide has a different amino acid sequence from another reference polypeptide. Generally, the differences are limited such that the sequences of the reference polypeptide and the variant are generally very similar and identical in many regions. The variant polypeptide and the reference polypeptide may have different amino acid sequences due to one or more substitutions, additions, or deletions in any combination. The substituted or inserted amino acid residues may or may not be residues encoded by the genetic code. Variants of polynucleotides or polypeptides may be naturally occurring, such as allelic variants, or unknown naturally occurring variants. Polynucleotide and polypeptide variants that do not exist naturally may be produced by mutagenesis, direct synthesis, and other recombination methods known to those skilled in the art.

[0036] As used in this application, the expression "functional protein encoded by a retrotransposon" refers to a polypeptide that catalyzes the incorporation of an exogenous nucleic acid fragment into a target site (such as genome or extrachromosomal DNA).

[0037] As used in this application, the term “exogenous nucleic acid fragment” includes any gene of interest or any transferable gene or fragment thereof. In some non-limiting embodiments, the exogenous nucleic acid fragment is of a different origin from the host cell, for example, a nucleic acid sequence isolated from an organism other than the host cell, i.e., the exogenous nucleic acid fragment is exogenous to the host cell.

[0038] As used in this application, the terms “domain” and “functional domain” are interchangeable and refer to a structure of a biomolecule that contributes to a specific function of the biomolecule, and may include a continuous region (such as a continuous sequence) or a different discontinuous region (such as a discontinuous sequence) of the biomolecule. Examples of protein functional domains include, but are not limited to, DNA-binding domains, RNA-binding domains, reverse transcriptase functional domains, and nuclease functional domains.

[0039] As used in this application, the term “untranslated region” refers to nucleic acid sequences located at both ends of a transposable element and adjacent to the transposable nucleic acid sequence. Of these, the untranslated region located at the 5' end of the transposable nucleic acid sequence is called the 5' untranslated region, and the untranslated region located at the 3' end of the transposable nucleic acid sequence is called the 3' untranslated region. In some embodiments, RNA transcribed from an untranslated region can bind to a functional protein encoded by a specific retrotransposon.

[0040] As used in this application, the term “nucleic acid construct” is defined herein as a single-stranded or double-stranded nucleic acid molecule, and preferably refers to an artificially constructed nucleic acid molecule. Optionally, a nucleic acid construct further comprises one or more operablely linked regulatory sequences that can direct the expression of a coding sequence in a suitable host cell under suitable conditions. The term “expression” is understood to include, but is not limited to, any steps involved in the production of a protein or polypeptide, including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and secretion. The term “regulatory sequence” includes all components necessary or advantageous for the expression of polypeptides / proteins in this application. Each regulatory sequence may be naturally present in the nucleic acid sequence encoding a protein or polypeptide, or it may be exogenous. These regulatory sequences include, but are not limited to, leader sequences, polyadenylated sequences, propeptide sequences, promoters, signal sequences, and transcription terminators. At a minimum, a regulatory sequence should include promoters for transcription and translation, as well as start and end signals. Regulatory sequences having linkers may be provided for the purpose of introducing the regulatory sequence into a specific restriction site for linking the regulatory sequence to the coding region of the nucleic acid sequence encoding a protein or polypeptide.

[0041] As used in this application, the term “promoter” refers to a polynucleotide sequence capable of controlling the transcription of a coding sequence. A promoter sequence includes a sequence specific enough to enable RNA polymerase to recognize, bind to, and initiate transcription. Furthermore, a promoter sequence may optionally include sequences that modulate the recognition, binding, and transcription initiation activity of RNA polymerase in the nucleic acid construct provided in this application. A promoter may affect the transcription of genes located on the same nucleic acid molecule as the promoter or genes located on a different nucleic acid molecule than the promoter.

[0042] As used in this application, the term “host cell” includes, but is not limited to, animal cells, plant cells, algal cells, fungal cells, yeast cells, or bacterial cells. The term includes offspring of the original cell into which an exogenous nucleic acid fragment has been introduced. An example host cell is human embryonic kidney cell HEK293T. It is understood that, due to natural, accidental, or intentional mutations, offspring of a single parental cell may not necessarily be morphologically or with respect to the original parent in terms of genomic or total DNA complement.

[0043] As used in this application, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid molecule to which it is ligated. Examples of vectors include, but are not limited to, plasmids, viruses, bacteria, phages, and insertable DNA fragments. The term “plasmid” refers to a circular double-stranded DNA that can receive an exogenous nucleic acid fragment and replicate in prokaryotic or eukaryotic cells.

[0044] Functional proteins encoded by retrotransposons This application provides a non-LTR retrotransposon system and its use. According to one embodiment of this application, an isolated functional protein encoded by a retrotransposon can be provided, having a functional protein sequence selected from (i) below, or a variant sequence of the aforementioned functional protein having the functional protein activity of (ii) to (iv): (i) at least one amino acid sequence represented by any one of SEQ ID NOs: 1 to 94; (ii) at least one sequence having deletion, substitution, insertion, or mutation of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids of the amino acid sequence represented by any one of SEQ ID NOs: 1 to 94; (iii) at least one amino acid sequence having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence represented by any one of SEQ ID NOs: 1 to 94; and (iv) at least one sequence obtained by further fusing the amino acid sequence represented by any one of SEQ ID NOs: 1 to 94 with another sequence.

[0045] According to one embodiment of this application, an isolated functional protein encoded by a retrotransposon, wherein the functional protein is of formula (I): C (X1) a C (X2) b H (X3) c H (I), A functional protein can be provided that contains the amino acid sequence shown in (of which a, b, and c are the number of amino acids, C is cysteine, H is histidine, (X1) is any amino acid, a is 1, 2, 3, or 4, (X2) is any amino acid, b is 11, 12, 13, 14, 15, 16, or 17, (X3) is any amino acid, and c is 4).

[0046] According to one embodiment of this application, an isolated functional protein encoded by a retrotransposon, wherein the functional protein is of formula (II): G (X4) d QGD (X5) e S (X6) f F (X7) g D (II), A functional protein can be provided that contains the amino acid sequence shown below (of which d, e, f, and g are the number of amino acids, D is aspartic acid, F is phenylalanine, G is glycine, Q is glutamine, S is serine, (X4) is any amino acid, d is 2, (X5) is any amino acid, e is 2, (X6) is any amino acid, f is 3, (X7) is any amino acid, and g is 30, 31, 32, 33, 34, 35, or 36).

[0047] According to one embodiment of this application, an isolated functional protein wherein the functional protein is of formula (III): C (X8) h C (X9) i E (X 10 ) j H (X 11 ) k C (X12 ) l RH (X 13 ) m PD (X 14 ) n (X 15 ) (X 16 ) o K (X 17 ) p Y (III), (Of these, h, i, j, k, l, m, n, o and p are the number of amino acids, C is cysteine, D is aspartic acid, E is glutamic acid, H is histidine, K is lysine, R is arginine, P is proline, Y is tyrosine, (X8) is any amino acid, h is 2, 3 or 4, (X9) is any amino acid, i is 3, 4, 5, 6, 7, 8, 9 or 10, (X 10 ) is any amino acid, j is 3, and (X 11 ) is any amino acid, k is 4, and (X 12 ) is any amino acid, l is 9, and (X 13 ) is any amino acid, m is 31, 32, 33, 34, 35 or 36, (X 14 ) is any amino acid, where n is 11, 12, 13, 14, 15, 16, 17, 18 or 19, (X 15 ) is aspartic acid or glutamic acid, (X 16 ) is any amino acid, o is 15, 16, 17, 18 or 19, (X 17 A functional protein can be provided that contains the amino acid sequence shown in (where p is any amino acid and p is 3).

[0048] According to one embodiment of this application, a functional protein is provided which is an isolated functional protein encoded by a retrotransposon and comprises at least two of the amino acid sequences shown in formulas (I), (II), and (III).

[0049] According to one embodiment of this application, an isolated functional protein encoded by a retrotransposon is provided, comprising the amino acid sequences shown in formulas (I), (II), and (III).

[0050] In some embodiments, the retrotransposon is a non-LTR (non-long terminal repeat) retrotransposon. In some embodiments, the non-LTR retrotransposon is a restriction enzyme-like nuclease (RLE) retrotransposon. In some embodiments, the retrotransposon is a retrotransposon of the R2 family.

[0051] In some embodiments, the functional protein includes a DNA-binding domain that can bind to a specific region in the cell's genome, preferably the DNA-binding domain can bind to a gene encoding 28s rRNA. In some non-limiting examples, the functional protein takes a 28s rRNA gene as its target site. In some non-limiting examples, the functional protein takes a non-28s rRNA gene as its target site.

[0052] In some embodiments, the functional protein further comprises an RNA-binding domain, a reverse transcriptase functional domain, and / or a nuclease functional domain. In some non-limiting embodiments, the RNA-binding domain of the functional protein associates with RNA transcribed by the retrotransposon to form an RNP (ribonucleoprotein). In some non-limiting embodiments, the functional protein comprises a nuclease functional domain and / or a reverse transcriptase functional domain. The nuclease functional domain forms a single-strand nick at the DNA target site, and then uses the 3' end of the DNA exposed by this cleavage to prime the reverse transcription of the retrotransposon, with the transcribed RNA serving as a template for reverse transcription. Finally, insertion into the target site is achieved by "copy and paste".

[0053] In some embodiments, the species from which the functional protein is derived may be arthropods or chordates. In some embodiments, the species from which the functional protein is derived may be insects, Actinopterygii, Chondrichthyes, Testudines, Lepidosauria, or birds. In some embodiments, the species from which the functional protein is derived are Accipiter gentilis, Actinemys marmorata, Agelaius tricolor, Agrochola macilenta, Anagrus nilaparvatae, Andrena haemorrhoa, Anoplius nigerrimus, Artemisiospiza belli, Asobara japonica, Athalia rosae, Blastobasis lacticolella, and Bombus holtorum. Bombus hortorum, Bombus hypnorum, Bombus pratorum, Bombus vancouverensis, Brenthis ino, Cardiocondyla obscurior, Cerceris rybyensis, Clusia tigrina, Colaptes auratus, Crematogaster levior, Crotalus tigris, Cuora mccordi, Dinocampus cocchinellaecoccinellae), Dolichovespula saxonica, Drosophila albomicans, Drosophila saltans, Ennomos quercinarius, Eopsaltria australis, Eristalis pertinax, Erithacus rubecula, Euphyes dion, Heliconius hecale, Hesperophylax magnus, Ichneumon xanthorius, Jera trixpidata tricuspidata), Junonia litoralis, Lasioglossum baleicum, Lasioglossum lativentre, Lasioglossum morio, Leptopilina heterotoma, Lysandra coridon, Marasmarcha lunaedactyla, Mimumesa dahlbomi, Muschampia plurimacula, Neodiprion fabricii, Neodiprion pinetum, Nephrotoma flavescens flavescens), Nomada fabriciana, Nylanderia fulva, Nymphalis io, Papilio machaon, Pararge aegeriaaegeria), Podocnemis expansa, Poecilia wingei, Prinia subflava, Scaptomyza hsui, Schistocerca americana, Seladonia tumulorum, Sphecodes monilicornis, Triplophysa tibetana, Trypoxylus dichotomus, Urbanus tucuti, Venturia canescens, Vespa This includes crabro, Zaprionus camerounensis, and Zaprionus kolodkinae.

[0054] According to one embodiment of this application, nucleic acids encoding the functional protein described in this application can be provided.

[0055] Nucleic acid construct According to one embodiment of this application, a nucleic acid set can be provided, the nucleic acid set comprising a 5' untranslated region, the 5' untranslated region comprising at least one of the nucleotide sequences shown in SEQ ID NOs. 95 to 188.

[0056] According to one embodiment of this application, a nucleic acid set can be provided, the nucleic acid set comprising a 3' untranslated region, the 3' untranslated region comprising at least one nucleotide sequence shown in SEQ ID NOs. 189 to 282.

[0057] According to one embodiment of this application, a nucleic acid set can be provided, the nucleic acid set comprising a 5' untranslated region and a 3' untranslated region, wherein the 5' untranslated region comprises a nucleotide sequence or a variant thereof shown in any one of SEQ ID NOs. 95 to 188, and the 3' untranslated region comprises a nucleotide sequence or a variant thereof shown in any one of SEQ ID NOs. 189 to 282, and RNA transcribed from the nucleic acid set can bind to a functional protein encoded by a specific retrotransposon. In some embodiments, RNA transcribed from the untranslated region can recognize and bind to the RNA-binding domain of a functional protein encoded by a specific retrotransposon.

[0058] According to one embodiment of this application, a nucleic acid construct can be provided, the nucleic acid construct comprising nucleic acids encoding the functional proteins and / or nucleic acid sets described in this application. In some embodiments, the nucleic acid construct further comprises exogenous nucleic acid fragments. In some embodiments, the exogenous nucleic acid fragments are operably inserted into the nucleic acid construct via polyclonal insertion sites, and there may be one or more exogenous nucleic acid fragments, which may be the same or different. In some embodiments, the exogenous nucleic acid fragments comprise any gene of interest or any transferable gene, preferably comprising genes for native functional proteins, artificial chimeric genes, and / or non-coding RNA genes. In some embodiments, the non-coding RNA genes comprise a variety of RNAs with known functions and RNAs with unknown functions, such as rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA). In some embodiments, the artificial chimeric gene comprises genes for chimeric antigen receptors. In some embodiments, the genes for the innate functional protein include a fluorescence-based reporter gene, a luciferase gene, and / or an antibiotic resistance gene. In some non-limiting embodiments, the exogenous nucleic acid fragment may further include a series of primers for polymerase chain reaction to facilitate screening of retrotransposition activity.

[0059] In some embodiments, the nucleic acid construct further optionally includes a homologous sequence having 100% identity with at least 10 nucleotides of a specific region in the cell genome, preferably further optionally including a homologous sequence having 100% identity with at least 10 nucleotides of a gene encoding 28s rRNA. In some non-limiting embodiments, a portion of the homologous sequence is identical to a target site to facilitate the initiation of the reverse transcription process. In some non-limiting embodiments, the nucleic acid construct includes the 5' untranslated region and the 3' untranslated region described in this application, with the homologous sequence upstream of the 5' untranslated region and / or downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequence is upstream of the 5' untranslated region. In some non-limiting embodiments, the homologous sequence is downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequence is upstream of the 5' untranslated region and downstream of the 3' untranslated region.

[0060] In some embodiments, the nucleic acid construct further comprises a promoter and a poly(A) sequence. The promoter may be any suitable promoter sequence, i.e., a nucleic acid sequence that can be recognized by a host cell expressing an exogenous nucleic acid fragment. The promoter sequence includes a transcriptional regulatory sequence that mediates the expression of a protein or polypeptide. The promoter may be any nucleic acid sequence (including mutant, truncated, and heterozygous promoters) that has transcriptional activity in a selected host cell and may originate from a gene encoding an extracellular or intracellular protein or polypeptide homologous or heterologous to the host cell. In some embodiments, the promoter may include CMV, EF1a, SV40, PGK, UbC, human beta-actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL. Poly(A) tailing signal arrays and various shortened forms of poly(A) tailing signals, which are well known in the art, can be used in this application.

[0061] In some embodiments, the nucleic acid construct further includes an optional transcription termination sequence (i.e., a sequence recognized by a host cell to terminate transcription) for controlling the expression of an exogenous nucleic acid fragment. Any terminator that functions in a selected host cell may be used in this application.

[0062] Optionally, the nucleic acid construct may further include a suitable leader sequence (i.e., an untranslated region in mRNA important for translation in the host cell) for controlling the expression of an exogenous nucleic acid fragment. The leader sequence is operably ligated to the 5' end of the nucleic acid sequence encoding the polypeptide. Any leader sequence that is functional in selected host cells may be used in this application.

[0063] Optionally, the nucleic acid construct may further include a propeptide coding region for controlling the expression of an exogenous nucleic acid fragment, the propeptide coding region encoding an amino acid sequence located at the amino terminus of the polypeptide. The resulting polypeptide is called a zymogen or propolypeptide. Propolypeptides are typically inactive and can be converted to mature, active polypeptides by catalytic or autocatalytic cleavage of the propeptide from the propolypeptide.

[0064] Optionally, the nucleic acid construct may further include a regulatory sequence that can regulate the expression of an exogenous nucleic acid fragment according to the growth conditions of the host cell. An example of a regulatory sequence is a system that turns gene expression on or off in response to chemical or physical stimuli, including the presence of a regulatory compound. Another example of a regulatory sequence is one that enables gene amplification. In these cases, the exogenous nucleic acid fragment should be operably ligated to the regulatory sequence.

[0065] Retrotransposition composition According to one embodiment of the present application, a composition may be provided which comprises a functional protein or functional fragment thereof encoded by an R2 family retrotransposon, or a nucleic acid encoding a functional protein or functional fragment thereof, wherein the functional protein or functional fragment thereof has the function of catalyzing the insertion of an exogenous nucleic acid fragment into the genome of a cell; and a nucleic acid set wherein the RNA transcribed by the nucleic acid set can be recognized by a functional protein or functional fragment thereof encoded by a particular retrotransposon.

[0066] In some embodiments, the composition is selected from at least one of the following groups (1) to (95), where any one of the following groups (1) to (94) includes: functional protein-associated sequences and nucleic acid sets, (1) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 95, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 189; (2) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 96, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 190; (3) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 97, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 191; (4) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 98, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 192; (5) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 99, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 193; (6) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 100, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 194; (7) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 101, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 195; (8) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 102, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 196; (9) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 103, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 197; (10) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 104, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 198; (11) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 105, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 199; (12) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 106, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 200; (13) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 107, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 201; (14) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 108, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 202; (15) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 109, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 203; (16) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 110, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 204; (17) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 111, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 205; (18) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 112, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 206; (19) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 113, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 207; (20) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 114, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 208; (21) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 115, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 209; (22) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 116, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 210; (23) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 117, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 211; (24) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 118, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 212; (25) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 119, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 213; (26) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 120, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 214; (27) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 121, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 215; (28) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 122, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 216; (29) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 123, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 217; (30) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 124, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 218; (31) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 125, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 219; (32) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 126, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 220; (33) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 127, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 221; (34) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 128, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 222; (35) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 129, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 223; (36) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 130, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 224; (37) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 131, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 225; (38) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 132, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 226; (39) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 133, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 227; (40) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 134, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 228; (41) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 135, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 229; (42) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 136, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 230; (43) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 137, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 231; (44) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 138, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 232; (45) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 139, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 233; (46) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 140, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 234; (47) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 141, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 235; (48) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 142, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 236; (49) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 143, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 237; (50) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 144, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 238; (51) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 145, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 239; (52) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 146, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 240; (53) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 147, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 241; (54) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 148, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 242; (55) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 149, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 243; (56) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 150, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 244; (57) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 151, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 245; (58) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 152, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 246; (59) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 153, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 247; (60) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 154, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 248; (61) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 155, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 249; (62) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 156, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 250; (63) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 157, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 251; (64) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 158, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 252; (65) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 159, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 253; (66) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 160, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 254; (67) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 161, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 255; (68) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 162, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 256; (69) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 163, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 257; (70) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 164, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 258; (71) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 165, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 259; (72) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 166, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 260; (73) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 167, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 261; (74) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 168, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 262; (75) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 169, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 263; (76) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 170, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 264; (77) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 171, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 265; (78) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 172, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 266; (79) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 173, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 267; (80) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 174, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 268; (81) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 175, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 269; (82) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 176, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 270; (83) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 177, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 271; (84) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 178, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 272; (85) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 179, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 273; (86) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 180, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 274; (87) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 181, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 275; (88) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 182, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 276; (89) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 183, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 277; (90) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 184, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 278; (91) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 185, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 279; (92) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 186, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 280; (93) The functional protein-related sequence is an amino acid sequence or nucleic acid encoding an amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 187, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 281; (94) The functional protein-related sequence is an amino acid sequence or a nucleic acid encoding an amino acid sequence, wherein the nucleic acid set comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 188, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 282; or (95) A mutant of any of the above groups (1) to (94), Here, the functional protein-related sequence is the amino acid sequence of a variant of the functional protein of each group or a nucleic acid sequence encoding the variant, and the variant has a variant sequence of the functional protein having functional protein activity selected from (i) to (iii) below: (i) at least one sequence having deletion, substitution, insertion, or mutation of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids of the amino acid sequence of the functional protein of each group; (ii) at least one amino acid sequence having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence shown in any one of sequence numbers 1 to 94; and (iii) at least one sequence obtained by further fusing the amino acid sequence shown in any one of sequence numbers 1 to 94 with another sequence.

[0067] In some embodiments, the nucleic acid set further comprises exogenous nucleic acid fragments. In some embodiments, the exogenous nucleic acid fragments are operably inserted into the nucleic acid construct via polyclonal insertion sites, and there may be one or more exogenous nucleic acid fragments, which may be the same or different. In some embodiments, the exogenous nucleic acid fragments comprise any gene of interest or any transferable gene, preferably comprising genes for native functional proteins, artificial chimeric genes, and / or non-coding RNA genes. In some embodiments, the non-coding RNA genes comprise a variety of RNAs with known functions and RNAs with unknown functions, such as rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA). In some embodiments, the artificial chimeric gene comprises a gene for a chimeric antigen receptor. In some embodiments, the genes for native functional proteins comprise a fluorescence-based reporter gene, a luciferase gene, and / or a resistance gene. In some non-limiting embodiments, the exogenous nucleic acid fragment may further include a series of primers for polymerase chain reaction to facilitate screening for retrotransposition activity.

[0068] In some embodiments, the nucleic acid set further optionally includes homologous sequences having 100% identity with at least 10 nucleotides of a specific region in the cell genome, preferably further optionally including homologous sequences having 100% identity with at least 10 nucleotides of a gene encoding 28s rRNA. In some non-limiting embodiments, a portion of the homologous sequences is identical to a target site to facilitate the initiation of the reverse transcription process. In some non-limiting embodiments, the nucleic acid construct includes the 5' untranslated region and the 3' untranslated region described in this application, with the homologous sequences upstream of the 5' untranslated region and / or downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequences are located upstream of the 5' untranslated region. In some non-limiting embodiments, the homologous sequences are located downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequences are located upstream of the 5' untranslated region and downstream of the 3' untranslated region.

[0069] In some embodiments, the nucleic acid and / or nucleic acid set further comprises a promoter and a poly(A) sequence. The promoter may be any suitable promoter sequence, i.e., a nucleic acid sequence that can be recognized by a host cell expressing the nucleic acid sequence. The promoter sequence includes a transcriptional regulatory sequence that mediates the expression of a protein or polypeptide. The promoter may be any nucleic acid sequence (including mutant, truncated, and heterozygous promoters) that has transcriptional activity in selected host cells and may originate from a gene encoding an extracellular or intracellular protein or polypeptide homologous or heterologous to the host cell. In some embodiments, the promoter may include CMV, EF1a, SV40, PGK, UbC, human beta-actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL. Poly(A) tailing signal arrays and various shortened forms of poly(A) tailing signals, which are well known in the art, can be used in this application.

[0070] In some embodiments, the nucleic acid encoding an amino acid sequence and / or a set of nucleic acids further comprises an optional transcription termination sequence, i.e., a sequence recognized by a host cell to terminate transcription. The termination sequence is operably ligated to the 3' end of the nucleic acid sequence encoding a protein or polypeptide. Any terminator that functions in a selected host cell can be used in this application.

[0071] Optionally, a nucleic acid encoding an amino acid sequence and / or a set of nucleic acids may further include a suitable leader sequence, i.e., an untranslated region in mRNA that is important for translation in the host cell. The leader sequence is operably ligated to the 5' end of the nucleic acid sequence encoding the polypeptide. Any leader sequence that is functional in selected host cells may be used in this application.

[0072] Optionally, a nucleic acid encoding an amino acid sequence and / or a set of nucleic acids may further include a propeptide coding region that encodes an amino acid sequence located at the amino terminus of the polypeptide. The resulting polypeptide is called a zymogen or propolypeptide. Propolypeptides are typically inactive and can be converted to mature, active polypeptides by catalytic or autocatalytic cleavage of the propeptide from the propolypeptide.

[0073] Optionally, nucleic acids encoding amino acid sequences and / or sets of nucleic acids may further include regulatory sequences that can regulate polypeptide expression according to host cell growth conditions. Examples of regulatory sequences include systems that turn gene expression on or off in response to chemical or physical stimuli, including in the presence of regulatory compounds. Other examples of regulatory sequences are those that enable gene amplification. In these examples, the nucleic acid sequence encoding a protein or polypeptide should be operably ligated to the regulatory sequence.

[0074] Recombinant vectors, recombinant host cells, and kits According to one embodiment of this application, a recombinant vector can be provided, comprising a nucleic acid encoding a functional protein described herein, a nucleic acid described herein, a nucleic acid set described herein, a nucleic acid construct described herein, or a composition described herein. The recombinant vector may be any suitable vector. In some embodiments, the recombinant vector may be, but is not limited to, a recombinant cloning vector, a recombinant eukaryotic expression plasmid, or a recombinant viral vector. In some embodiments, the recombinant eukaryotic expression plasmid may include pcDNA3.1, pCMV, pUC18, pUC19, pUC57, pBAD, pET, pENTR, pGenlenti, or pAAV. In some embodiments, the recombinant viral vector may include a recombinant adenovirus vector, a recombinant adeno-associated virus vector, a recombinant retrovirus vector, a recombinant herpes simplex virus vector, or a recombinant vaccinia virus vector. The recombinant vectors of this application may be constructed using methods well known in the art. For example, depending on the restriction sites contained in the backbone vector used, appropriate restriction sites may be added to both ends of the nucleic acid construct of this application and then loaded into the backbone vector.

[0075] According to one embodiment of this application, a recombinant host cell can be provided, comprising a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid described in this application, a nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, or a recombinant vector described in this application. The recombinant host cell may be any host cell to which a retrotransposon can be used. In some embodiments, the recombinant host cell may be, but is not limited to, an animal cell, a plant cell, an algal cell, a fungal cell, a yeast cell, or a bacterial cell. In some embodiments, the animal cell may include a mammalian cell. In some embodiments, mammalian cells include primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, pigment cells, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal cell lines), and cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW62). This includes (0, HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, and Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, and D3) and their differentiated cells, or induced pluripotent stem cell lines and their differentiated cells.

[0076] According to one embodiment of this application, a kit can be provided which comprises a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid described in this application, a nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, a recombinant vector described in this application, or a recombinant host cell described in this application.

[0077] Method and Use The retrotransposon-based tools and methods for large fragment gene insertion and integration provided in this application can be applied to many fields, including gene therapy, crop breeding, model animal engineering, and industrial microbial engineering. In particular, in the fields of gene therapy and cell therapy, the tools and methods can be applied to gene writing therapy, which is of great importance for the treatment of genetic diseases requiring long fragment gene correction.

[0078] According to one embodiment of this application, a method can be provided for introducing an exogenous nucleic acid fragment into the genome of a host cell, the method comprising delivering a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid described in this application, a nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, or a recombinant vector described in this application to a host cell.

[0079] According to one embodiment of the present application, a method for editing the genome of a host cell can be provided, the method comprising delivering a functional protein described in the present application, a nucleic acid encoding a functional protein described in the present application, a nucleic acid described in the present application, a nucleic acid set described in the present application, a nucleic acid construct described in the present application, a composition described in the present application, or a recombinant vector described in the present application to a host cell.

[0080] According to one embodiment of the present application, a method can be provided for obtaining a host cell containing an exogenous nucleic acid fragment in its genome, the method comprising delivering a functional protein described in the present application, a nucleic acid encoding a functional protein described in the present application, a nucleic acid described in the present application, a nucleic acid set described in the present application, a nucleic acid construct described in the present application, a composition described in the present application, or a recombinant vector described in the present application to a host cell.

[0081] The method of delivery to host cells can be any suitable method. In some embodiments, delivery methods include, but are not limited to, cationic liposome delivery, lipoid nanoparticle delivery, cationic polymer delivery, vesicle-exosome delivery, gold nanoparticle delivery, polypeptide and protein delivery, retroviral delivery, lentiviral delivery, adenovirus delivery, adeno-associated virus delivery, electroporation delivery, Agrobacterium infection delivery, or gene gun delivery. Methods of cell transfection and culture are routine in the art, and suitable transfection and culture methods can be selected according to different cell types.

[0082] The host cell can be any host cell from which retrotransposons can be used. In some embodiments, the host cell may be, but is not limited to, animal cells, plant cells, algal cells, fungal cells, yeast cells, or bacterial cells. In some embodiments, the host cell may be a mammalian cell. In some embodiments, the host cell may be primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, pigment cells, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / xylem cell lines), or cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW620). This includes HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, and Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, and D3) and their differentiated cells, or induced pluripotent stem cell lines and their differentiated cells.

[0083] According to one embodiment of this application, the use of a functional protein described in this application, a nucleic acid encoding a functional protein described in this application, a nucleic acid described in this application, a nucleic acid set described in this application, a nucleic acid construct described in this application, a composition described in this application, a recombinant vector described in this application, or a recombinant host cell described in this application can be provided for introducing an exogenous nucleic acid fragment into the genome of a host cell. The host cell may be any host cell on which a retrotransposon can be used. In some embodiments, the host cell may be, but is not limited to, an animal cell, a plant cell, an algal cell, a fungal cell, a yeast cell, or a bacterial cell. In some embodiments, the host cell may include a mammalian cell. In some embodiments, the host cells are primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, pigment cells, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal cell lines), cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW620) This includes HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, and Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, and D3) and their differentiated cells, or induced pluripotent stem cell lines and their differentiated cells.

[0084] According to one embodiment of this application, it is possible to provide the use of the functional protein described in this application, the nucleic acid encoding the functional protein described in this application, the nucleic acid described in this application, the nucleic acid set described in this application, the nucleic acid construct described in this application, the composition described in this application, the recombinant vector described in this application, or the recombinant host cell described in this application for gene therapy, cell therapy, genome research, or induction and post-induction differentiation of stem cells.

[0085] The various embodiments and preferences of this application described above can be combined with one another (inso that they are not inherently contradictory to each other and are suitable for use in this application), and the various embodiments formed by such combinations shall be considered part of this application. [Examples]

[0086] The exemplary embodiments of this application are described below in conjunction with the accompanying drawings and include various details of the examples of this application for the sake of ease of understanding. It should be understood that these are illustrative only and are not intended to limit the scope of protection of this application. The scope of protection of this application is defined solely by the claims. Therefore, those skilled in the art should be aware that various changes and modifications can be made to the examples described herein without departing from the scope of this application. Similarly, for the sake of clarity and brevity, descriptions of well-known functions and structures are omitted in the following description.

[0087] Unless otherwise specified, the reagents and equipment used in the following examples are commercially available conventional products. Unless otherwise specified, experiments are conducted under conventional conditions or conditions recommended by the manufacturer.

[0088] Example 1: Establishment of a screening system for retrotransposon activity The inventors have designed a high-throughput screening system for detecting the insertion activity of different retrotransposons of this application at specific sites in the 28s rRNA gene of the human genome.

[0089] The order of elements within the vector is shown in Figure 1, and a detailed explanation is as follows. First, we commissioned BGI TECH SOLUTIONS (BEIJING LIUHE) CO., LIMITED to synthesize the corresponding DNA sequence and construct a retrotransposon plasmid. A schematic diagram of an example of the retrotransposon construct is shown in Figure 1, which has a 5' untranslated region (5'UTR), a DNA sequence encoding the amino acid sequence of a functional protein optimized by human codons (ORF), and a 3' untranslated region (3'UTR) from the 5' end to the 3' end. The synthesized sequence was cloned into the plasmid vector pCDNA3.1, which already contained the CMV promoter element and poly(A) element, and the retrotransposon was transcribed in eukaryotic cells under the control of the CMV promoter, followed by translation into a functional protein.

[0090] Next, based on the functional characteristics of the R2 retrotransposon, the inventors added sequences (R105 and R100 in Figure 1) to both sides of the 5'UTR and 3'UTR of the R2 retrotransposon. The added sequences are homologous to the sequences on either side of the insertion site of the human 28S rRNA gene and were intended to enhance the integrity of the retrotransposition product (Eickbush DG, Luan DD, Eickbush TH. Integration of Bombyx mori R2 sequences into the 28S ribosomal RNA genes of Drosophila melanogaster. Mol Cell Biol. 2000;20(1):213-23.). These two homologous sequences are transcribed together into RNA during the retrotransposon transcription process.

[0091] To detect the retrotransposition activity of all retrotransposons using a unified method, the inventors inserted a consensus sequence (universal sequence) between the ORF region encoding a functional protein and the 3'UTR, and included a PCR primer F1 (5'-TGTGCCGAGGCTCAGGCACGCTC-3', having the sequence shown in SEQ ID NO: 301) with high specificity and amplification efficiency in this sequence. After the retrotransposition process was completed and R2 was inserted into the 28s rRNA gene, genomic DNA was extracted and used as a template for amplifying the junction of the 28s rRNA gene with the retrotransposon using F1 and a reverse primer a1 (5'-GGCCTCCCACTTATTCTACACC-3', having the sequence shown in SEQ ID NO: 302) approximately 200 bp downstream of the insertion site. PCR products of different lengths would be amplified for different R2 samples due to differences in the length of their 3'UTR sequences. The products were further analyzed by Sanger sequencing to confirm whether directional insertion at the 28s rRNA locus was achieved.

[0092] [Table 1-1]

[0093] [Table 1-2]

[0094] [Table 1-3]

[0095] Example 2: Retrotransposition Activity Assay 2.1 Cell treatment (Day 0): After culturing commercially available HEK293T cells until the logarithmic growth phase, they were digested with 0.25% trypsin (Thermo) to disperse into single cells and then placed in 96-well cell culture plates pre-coated with PDL (Sigma) at a cell concentration of 1 × 10⁶. 4The cells were added to each well and incubated overnight at 37°C under 5% CO2 conditions.

[0096] 2.2 Cell transfection (Day 1): 200 ng of each retrotransposon plasmid constructed in Example 1 was mixed with the transfection reagent Lipofectamine 2000 (Thermo) in a ratio of transfection plasmid mass (μg): transfection reagent volume (μL) = 1:2, and allowed to stand at room temperature for 15 minutes to form a transfection complex. The transfection complex was transferred to a cell culture plate and incubated with cells, and two parallel tests were performed for each sample to be screened.

[0097] 2.3 Cell collection and genome extraction (Day 4) 72 hours after transfection, the cells were digested and dispersed into single cells using 0.25% trypsin (Thermo). The cells were collected by centrifugation, and genomic DNA was extracted using a genome extraction kit (BEIJING BIOTEKE BIOTECHNOLOGY CO.,LTD.,DP1202). The concentration of genomic DNA was measured using a nanodrop one ultramicrovolume spectrophotometer (Thermo).

[0098] 2.4 Detection Genomic DNA from all samples was subjected to PCR analysis according to the conditions in Tables 2 and 3, and the PCR products were detected by 1% agarose gel electrophoresis.

[0099] [Table 2]

[0100] [Table 3]

[0101] 2.5 Detection Results The results for retrotransposition activity are shown in Figure 2. Different lanes in Figure 2 correspond to different retrotransposons numbered 1 to 100 in Table 1. The results show that the lanes corresponding to most retrotransposons show a single band (e.g., retrotransposons numbered 1-5, 7-12, 14-17, 19-32, 34-92, and 95-100 in Table 1), indicating that these functional proteins encoded by retrotransposons have specific retrotransposition activity. However, the lanes corresponding to six retrotransposons do not have a band (e.g., retrotransposons numbered 6, 13, 18, 33, 93, and 94 in Table 1), indicating that these functional proteins encoded by retrotransposons do not have retrotransposition activity. Compared to these inactive retrotransposons (numbered 6, 13, 18, 33, 93, and 94 in Table 1), the other 94 retrotransposons exhibit better retrotransposition activity (numbered 1-5, 7-12, 14-17, 19-32, 34-92, and 95-100 in Table 1).

[0102] Example 3: Relative quantitative analysis of transposon activity Using the human single-copy gene 36b4 as an internal reference gene, we performed a relative quantitative analysis of the retrotransposition activity of several retrotransposons listed in Table 1. The method was the same as that described in Examples 1 and 2, with the only difference being: during the PCR process, an additional pair of PCR primers targeting the single-copy gene 36b4 was added simultaneously. 36b4-F:5'-CAGCAAGTGGGAAGGTGTAATCC-3' (The sequence is as shown in sequence number 303); 36b4-R:5'-CCCATTCTATCATCAACGGGTACAA-3' (the sequence is as shown in sequence number 304).

[0103] The results of retrotransposition activity are shown in Figure 3. The numbers on different lanes correspond to the corresponding numbered transposon vectors in Table 1. The detection results in Figure 3 were quantitatively analyzed using Image Lab version 6.1.0 (Bio-Rad Laboratories Inc) software. The calculation formula is: gray value of the R2 band / gray value of the 36b4 band. The quantitative analysis results are shown in Figure 4 and Table 4. The above results indicate that all functional proteins encoded by the retrotransposon of this application have constant retrotransposition activity, and some functional proteins have relatively high activity.

[0104] [Table 4]

[0105] Example 4: Detection of the 5' junction of a retrotransposon The method used to detect the 5' junction of the 28s rRNA gene with the retrotransposon shown in Table 1 (demonstrating the completeness of the retrotransposition event) is as follows: The method is the same as that described in Examples 1 and 2, with the only difference being: three reverse primers were designed at different positions of the transposon ORF region and the 5' UTR sequence (the sequences of the reverse primers are shown in Table 4), each paired with a forward primer (28s-up-F1, with the sequence shown in SEQ ID NO: 305) approximately 200 bp upstream of the insertion site of the 28s rRNA gene for PCR analysis. The completeness of the 5' terminal sequence of these samples after insertion was detected to preliminarily determine the retrotransposition completeness of different R2 samples.

[0106] The results of the retrotransposition integrity assay are shown in Figure 5. The numbers on the different lanes correspond to the corresponding numbered transposon vectors in Table 1. The three lanes for each vector correspond to the PCR bands obtained by pairing three reverse primers (having the sequences shown in Table 5) with a forward primer (28s-up-F1 having the sequence shown in SEQ ID NO: 305). The results indicate that the retrotransposition products obtained from functional proteins encoded by the retrotransposons of this application exhibit relatively good integrity.

[0107] [Table 5-1]

[0108] [Table 5-2]

[0109] [Table 5-3]

[0110] It should be noted that the above are merely preferred examples of this application and are not intended to limit it. Those skilled in the art can make various modifications and changes to this application. While specific embodiments have been described, it is not possible, or even currently foreseeable, that the applicant or those skilled in the art may have substitutions, modifications, changes, improvements, and substantial equivalents of the above embodiments. Therefore, the submitted and potentially modified claims are intended to cover all such substitutions, modifications, changes, improvements, and substantial equivalents. It is important to note that as the art evolves, many of the elements described herein may be replaced by equivalent elements appearing after this application.

Claims

1. Isolated functional proteins encoded by retrotransposons, comprising a functional protein sequence selected from (i) below or a variant sequence of the aforementioned functional protein having the functional protein activities of (ii) to (iv): (i) At least one amino acid sequence shown in any one of Sequence IDs 1 to 94; (ii) At least one sequence having one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, substitutions, insertions, or mutations of the amino acid sequence shown in any one of sequence numbers 1 to 94; (iii) at least one amino acid sequence having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence shown in any one of Sequence IDs 1 to 94; and (iv) At least one sequence obtained by further fusing the amino acid sequence shown in any one of sequence numbers 1 to 94 with another sequence.

2. An isolated functional protein encoded by a retrotransposon, wherein the functional protein is of formula (I): C (X １ ) ａ C (X ２ ) ｂ H (X ３ ) ｃ H (I)、 (In the formula, a, b, and c are the number of amino acids, C is cysteine, H is histidine, (X １ ) is any amino acid, and a is 1, 2, 3, or 4. (X ２ ) is any amino acid, and b is 11, 12, 13, 14, 15, 16, or 17. (X ３ ) is an arbitrary amino acid, and c is 4), an isolated functional protein comprising the amino acid sequence shown.

3. An isolated functional protein encoded by a retrotransposon, wherein the functional protein is of formula (II): (︸ ４ ) ｄ . . . . . . . ５ ) ｅ 3(︸ ６ ) ｆ ︦ ７ ) ｇ .(......)、 (In the formula, d, e, f, and g are the number of amino acids. D is aspartic acid, F is phenylalanine, G stands for glycine, Q is glutamine, S is serine, (X ４ ) is any amino acid, and d is 2, (X ５ ) is any amino acid, and e is 2, (X ６ ) is any amino acid, and f is 3, (X ７ An isolated functional protein containing the amino acid sequence shown in (where g is any amino acid, and g is 30, 31, 32, 33, 34, 35, or 36).

4. An isolated functional protein encoded by a retrotransposon, wherein the functional protein is of formula (III): C (X ８ ) ｈ C (X ９ ) ｉ E (X １０ ) ｊ H (X １１ ) ｋ C (X １２ ) ｌ R H (X １３ ) ｍ P D (X １４ ) ｎ (X １５ ) (X １６ ) ｏ K (X １７ ) ｐ Y (III)、 (In the formula, h, i, j, k, l, m, n, o, and p are the number of amino acids. C is cysteine, D is aspartic acid, E is glutamic acid, H is histidine, K is lysine, R is arginine, P is proline, Y is tyrosine, (X ８ ) is any amino acid, and h is 2, 3, or 4. (X ９ ) is any amino acid, where i is 3, 4, 5, 6, 7, 8, 9, or 10. (X １０ ) is any amino acid, and j is 3, (X １１ ) is any amino acid, and k is 4, (X １２ ) is any amino acid, and l is 9, (X １３ ) is any amino acid, and m is 31, 32, 33, 34, 35, or 36. (X １４ ) is any amino acid, where n is 11, 12, 13, 14, 15, 16, 17, 18, or 19. (X １５ ) is aspartic acid or glutamic acid, (X １６ ) is any amino acid, and o is 15, 16, 17, 18 or 19. (X １７ An isolated functional protein containing the amino acid sequence shown in (where p is any amino acid and p is 3).

5. An isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises at least two of the amino acid sequences shown in formulas (I), (II), and (III).

6. Isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises the amino acid sequences shown in formulas (I), (II), and (III).

7. The functional protein according to any one of claims 1 to 6, wherein the retrotransposon is a non-LTR (non-long terminal repeat) retrotransposon.

8. The functional protein according to claim 7, wherein the non-LTR retrotransposon is a restriction enzyme-like nuclease (RLE) retrotransposon.

9. The functional protein according to any one of claims 1 to 8, wherein the retrotransposon is an R2 family retrotransposon.

10. The functional protein according to any one of claims 1 to 9, wherein the functional protein comprises a DNA-binding domain that can bind to a specific region in the genome of a cell, and preferably the DNA-binding domain can bind to a gene encoding 28s rRNA.

11. The functional protein according to claim 10, wherein the functional protein further comprises an RNA-binding domain, a reverse transcriptase functional domain, and / or a nuclease functional domain.

12. The functional protein according to any one of claims 1 to 11, wherein the species from which the functional protein is derived includes arthropods or chordates.

13. The functional protein according to claim 12, wherein the species from which the functional protein is derived includes the class Insecta, subclass Actinopterygii, class Chondrichthyes, order Testudines, order Lepidosauria, or birds.

14. The species from which the aforementioned functional protein originates are Accipiter gentilis, Actinemys marmorata, Agelaius tricolor, Agrochola macilenta, Anagrus nilaparvatae, Andrena haemorrhoa, Anoplius nigerrimus, Artemisiospiza belli, and Asobara japonica. japonica), Athalia rosae, Blastobasis lacticolella, Bombus hortorum, Bombus hypnorum, Bombus pratorum, Bombus vancouverensis, Brenthis ino, Cardiocondyla obscurior, Cerceris rybyensis, Clusia tigrina tigrina), Colaptes auratus, Crematogaster levior, Crotalus tigris, Cuora maccordi, Dinocampus coccinellae, Dolichovespula saxonica, Drosophylla albomicans, Drosophylla saltans, Ennomos quaercinarius quercinaurius, Eopsaltria australisaustralis), Eristalis pertinax, Eritacus rubecula, Euphyes dion, Heliconius hecale, Hesperophylax magnus, Ichneumon xanthorius, Jera tricuspidata, Junonia litoralis, Lasioglossum baraicum baleicum), Lasioglossum lativentre, Lasioglossum morio, Leptopilina heterotoma, Lysandra coridon, Marasmarcha lunaedactyla, Mimumesa dahlbomi, Muchampia plurimacula, Neodiprion fabricii fabricii), Neodiprion pinetum, Nephrotoma flavescens, Nomada fabriciana, Nylanderia fulva, Nymphalis io, Papilio machaon, Pararge aegeria, Podocnemis expansa, Poecilia wingei, Prinia subhlava subflava, Scaptomyza hsui, Schistocerca americana, Seladonia tumrolumThe functional protein according to claim 13, comprising Tumulorum, Specodes monilicornis, Triplophysa tibetana, Trypoxylus dichotomus, Urbanus tucti, Venturia canescens, Vespa crabro, Zaprionus cameroonensis, or Zaprionus kolodkinae.

15. A nucleic acid wherein the nucleic acid encodes a functional protein according to any one of claims 1 to 14.

16. A nucleic acid set comprising a 5' untranslated region, wherein the 5' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs. 95 to 188.

17. A nucleic acid set comprising a 3' untranslated region, wherein the 3' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 189 to 282.

18. A nucleic acid set comprising a 5' untranslated region and a 3' untranslated region, wherein the 5' untranslated region comprises a nucleotide sequence represented by any one of SEQ ID NOs. 95 to 188 or a variant thereof, and the 3' untranslated region comprises a nucleotide sequence represented by any one of SEQ ID NOs. 189 to 282 or a variant thereof, wherein RNA transcribed from the nucleic acid set can bind to a functional protein encoded by a specific retrotransposon.

19. A nucleic acid construct comprising the nucleic acid described in claim 15 and / or the nucleic acid set described in any one of claims 16 to 18.

20. The nucleic acid construct according to claim 19, further comprising exogenous nucleic acid fragments, the exogenous nucleic acid fragments being operably inserted into the nucleic acid construct via polyclonal insertion sites, wherein there may be one or more exogenous nucleic acid fragments, and they may be the same or different.

21. The nucleic acid construct according to claim 20, wherein the exogenous nucleic acid fragment comprises any gene of interest or any transferable gene, preferably comprising a gene for a natural functional protein, an artificial chimeric gene, and / or a gene for a non-coding RNA.

22. The nucleic acid construct according to claim 21, wherein the genes of the natural functional protein include a fluorescence-based reporter gene, a luciferase gene, and / or an antibiotic resistance gene.

23. The nucleic acid construct according to claim 21, wherein the artificial chimeric gene includes a gene for a chimeric antigen receptor.

24. The nucleic acid construct according to claim 21, wherein the non-coding RNA gene comprises rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA).

25. The nucleic acid construct according to claim 19, wherein the nucleic acid construct further comprises a promoter and a poly(A) sequence.

26. The nucleic acid construct according to claim 25, wherein the promoter comprises CMV, EF1a, SV40, PGK, UbC, human beta-actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL.

27. A composition, wherein the composition is A functional protein or functional fragment thereof encoded by an R2 family retrotransposon, or a nucleic acid encoding the said functional protein or functional fragment thereof, wherein the said functional protein or functional fragment thereof has the function of catalyzing the insertion of an exogenous nucleic acid fragment into the cellular genome, and A composition comprising a nucleic acid set, wherein the RNA transcribed by the nucleic acid set can be recognized by a functional protein or a functional fragment thereof encoded by a specific retrotransposon.

28. The composition according to claim 27, wherein the composition is selected from at least one of the following groups (1) to (95), and any one of the following groups (1) to (94) comprises: functional protein-related sequences and nucleic acid sets, (1) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 1 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 95, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 189; (2) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 2 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 96, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 190; (3) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 3 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 97, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 191; (4) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 4 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 98, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 192; (5) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 5 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 99, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 193; (6) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 6 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 100, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 194; (7) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 7 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 101, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 195; (8) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 8 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 102, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 196; (9) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 9 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 103, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 197; (10) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 10 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 104, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 198; (11) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 11 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 105, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 199; (12) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 12 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 106, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 200; (13) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 13 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 107, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 201; (14) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 14 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 108, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 202; (15) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 15 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 109, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 203; (16) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 16 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 110, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 204; (17) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 17 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 111, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 205; (18) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 18 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 112, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 206; (19) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 19 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 113, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 207; (20) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 20 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 114, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 208; (21) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 21 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 115, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 209; (22) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 22 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 116, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 210; (23) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 23 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 117, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 211; (24) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 24 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 118, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 212; (25) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 25 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 119, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 213; (26) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 26 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 120, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 214; (27) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 27 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 121, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 215; (28) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 28 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 122, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 216; (29) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 29 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 123, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 217; (30) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 30 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 124, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 218; (31) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 31 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 125, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 219; (32) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 32 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 126, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 220; (33) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 33 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 127, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 221; (34) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 34 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 128, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 222; (35) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 35 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 129, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 223; (36) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 36 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 130, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 224; (37) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 37 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 131, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 225; (38) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 38 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 132, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 226; (39) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 39 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 133, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 227; (40) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 40 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 134, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 228; (41) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 41 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 135, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 229; (42) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 42 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 136, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 230; (43) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 43 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 137, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 231; (44) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 44 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 138, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 232; (45) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 45 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 139, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 233; (46) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 46 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 140, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 234; (47) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 47 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 141, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 235; (48) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 48 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 142, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 236; (49) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 49 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 143, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 237; (50) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 50 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 144, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 238; (51) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 51 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 145, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 239; (52) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 52 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 146, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 240; (53) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 53 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 147, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 241; (54) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 54 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 148, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 242; (55) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 55 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 149, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 243; (56) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 56 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 150, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 244; (57) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 57 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 151, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 245; (58) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 58 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 152, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 246; (59) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 59 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 153, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 247; (60) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 60 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 154, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 248; (61) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 61 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 155, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 249; (62) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 62 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 156, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 250; (63) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 63 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 157, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 251; (64) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 64 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 158, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 252; (65) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 65 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 159, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 253; (66) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 66 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 160, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 254; (67) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 67 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 161, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 255; (68) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 68 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 162, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 256; (69) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 69 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 163, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 257; (70) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 70 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 164, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 258; (71) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 71 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 165, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 259; (72) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 72 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 166, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 260; (73) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 73 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 167, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 261; (74) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 74 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 168, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 262; (75) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 75 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 169, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 263; (76) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 76 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 170, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 264; (77) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 77 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 171, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 265; (78) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 78 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 172, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 266; (79) The functional protein-related sequence is an amino acid sequence containing the sequence shown in Sequence ID No. 79 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 173, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in Sequence ID No. 267; (80) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 80 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 174, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 268; (81) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 81 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 175, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 269; (82) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 82 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 176, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 270; (83) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 83 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 177, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 271; (84) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 84 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 178, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 272; (85) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 85 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 179, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 273; (86) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 86 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 180, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 274; (87) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 87 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 181, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 275; (88) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 88 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 182, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 276; (89) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 89 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 183, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 277; (90) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 90 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 184, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 278; (91) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 91 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 185, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 279; (92) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 92 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 186, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 280; (93) The functional protein-related sequence is an amino acid sequence containing the sequence shown in SEQ ID NO: 93 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 187, and the 3' untranslated region being a nucleotide sequence containing the sequence shown in SEQ ID NO: 281; (94) The functional protein-related sequence is an amino acid sequence including the sequence shown in SEQ ID NO: 94 or a nucleic acid encoding the amino acid sequence, wherein the nucleic acid set includes a 5' untranslated region and a 3' untranslated region, the 5' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 188, and the 3' untranslated region being a nucleotide sequence including the sequence shown in SEQ ID NO: 282; or (95) A mutant of any of the above groups (1) to (94), Here, the functional protein-related sequence is the amino acid sequence of a variant of the functional protein in each group or the nucleic acid sequence encoding the variant, and the variant has a variant sequence of the functional protein having functional protein activity selected from (i) to (iii) below: (i) At least one sequence having one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, substitutions, insertions, or mutations in the amino acid sequence of the functional protein of each group; (ii) at least one amino acid sequence having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence shown in any one of Sequence IDs 1 to 94; and (iii) At least one sequence obtained by further fusing the amino acid sequence shown in any one of Sequence IDs 1 to 94 with another sequence.

29. The composition according to claim 27 or 28, wherein the nucleic acid and / or nucleic acid set further comprises a promoter and a poly(A) sequence.

30. The composition according to claim 29, wherein the promoter comprises CMV, EF1a, SV40, PGK, UbC, human beta-actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL.

31. The composition according to claim 27 or 28, wherein the nucleic acid set further comprises exogenous nucleic acid fragments.

32. The composition according to claim 31, wherein the exogenous nucleic acid fragments are operably inserted into the nucleic acid set via polyclonal insertion sites, and the exogenous nucleic acid fragments may be one or more, and may be the same or different.

33. The composition according to claim 32, wherein the exogenous nucleic acid fragment comprises any gene of interest or any transferable gene, preferably comprising a gene for a natural functional protein, an artificial chimeric gene, and / or a gene for a non-coding RNA.

34. The composition according to claim 33, wherein the genes for the natural functional protein include a fluorescence-based reporter gene, a luciferase gene, and / or a resistance gene.

35. The composition according to claim 33, wherein the artificial chimeric gene comprises a gene for a chimeric antigen receptor.

36. The composition according to claim 33, wherein the non-coding RNA gene comprises rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA).

37. A recombinant vector comprising a nucleic acid encoding a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, or a composition according to any one of claims 27 to 36.

38. The recombinant vector according to claim 37, wherein the recombinant vector comprises a recombinant cloning vector, a recombinant eukaryotic expression plasmid, or a recombinant viral vector.

39. The recombinant vector according to claim 38, wherein the recombinant eukaryotic expression plasmid comprises pcDNA3.1, pCMV, pUC18, pUC19, pUC57, pBAD, pET, pENTR, pGenlenti, or pAAV.

40. The recombinant vector according to claim 38, wherein the recombinant viral vector includes a recombinant adenovirus vector, a recombinant adeno-associated virus vector, a recombinant retrovirus vector, a recombinant herpes simplex virus vector, or a recombinant vaccinia virus vector.

41. Recombinant host cell, wherein the recombinant host cell comprises a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, or a recombinant vector according to any one of claims 37 to 40.

42. The recombinant host cell according to claim 41, wherein the recombinant host cell includes an animal cell, a plant cell, an algal cell, a fungal cell, a yeast cell, or a bacterial cell.

43. The recombinant host cell according to claim 42, wherein the animal cell includes a mammalian cell.

44. The mammalian cells include primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, pigment cells, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), and cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW620, HCT-15, HCT1). Recombinant host cells according to claim 43, comprising 16, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, and Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, and D3) and their differentiated cells, or induced pluripotent stem cell lines and their differentiated cells.

45. A method for introducing an exogenous nucleic acid fragment into the genome of a host cell, the method comprising delivering to the host cell a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, or a recombinant vector according to any one of claims 37 to 40.

46. A method for editing the genome of a host cell, the method comprising delivering to the host cell a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, or a recombinant vector according to any one of claims 37 to 40.

47. A method for obtaining a host cell containing an exogenous nucleic acid fragment in its genome, the method comprising delivering to the host cell a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, or a recombinant vector according to any one of claims 37 to 40.

48. The method according to any one of claims 45 to 47, wherein the delivery method includes cationic liposome delivery, lipoid nanoparticle delivery, cationic polymer delivery, vesicle-exosome delivery, gold nanoparticle delivery, polypeptide and protein delivery, retrovirus delivery, lentivirus delivery, adenovirus delivery, adeno-associated virus delivery, electroporation delivery, Agrobacterium infection delivery, or gene gun delivery.

49. The method according to any one of claims 45 to 47, wherein the host cell includes an animal cell, a plant cell, an algal cell, a fungal cell, a yeast cell, or a bacterial cell.

50. The method according to claim 49, wherein the host cell includes a mammalian cell.

51. The host cells include primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, pigment cells, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), and cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW620, HCT-15, HCT The method according to claim 50, comprising 156, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, and Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, and D3) and their differentiated cells, or induced pluripotent stem cell lines and their differentiated cells.

52. Use of a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, a recombinant vector according to any one of claims 37 to 40, or a recombinant host cell according to any one of claims 41 to 44 for introducing an exogenous nucleic acid fragment into the genome of a host cell.

53. The use according to claim 52, wherein the host cell includes an animal cell, a plant cell, an algal cell, a fungal cell, a yeast cell, or a bacterial cell.

54. The use according to claim 53, wherein the host cell includes a mammalian cell.

55. The host cells include primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, pigment cells, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), and cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW620, HCT-15, HCT The use according to claim 54, comprising 156, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, and Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, and D3) and their differentiated cells, or induced pluripotent stem cell lines and their differentiated cells.

56. Use of a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, a recombinant vector according to any one of claims 37 to 40, or a recombinant host cell according to any one of claims 41 to 44 for preparing a drug or formulation for gene therapy, cell therapy, genome research, or stem cell induction and post-induction differentiation.

57. A kit comprising a functional protein according to any one of claims 1 to 14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16 to 18, a nucleic acid construct according to any one of claims 19 to 26, a composition according to any one of claims 27 to 36, a recombinant vector according to any one of claims 37 to 40, or a recombinant host cell according to any one of claims 41 to 44.

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