Gene transcription framework, vector system, genome sequence editing method and application

Abstract

The present invention provides a gene transcription framework, a vector system, a genome sequence editing method and its application. The gene transcription framework is based on the retrotransposition mechanism in eukaryotes. It can be mediated by DNA, RNA or RNP pathways, and includes one or more upstream sequences of the target site, a sequence to be inserted and a downstream sequence of the target site, one or more SINE (elements), one or more LINE (elements), one or more ORF1p coding sequences and / or one or more ORF2p coding sequences, and a transcription production sequence. According to the gene editing method, on the premise that foreign systems or substances are not introduced as much as possible and DNA double-strand breaks are not caused, the gene editing system is transferred to the cell nucleus or cytoplasm via the DNA, RNA or RNP pathway (RNA or RNP is transferred from the cytoplasm to the cell nucleus via ORF1p and / or ORF2p mediation), and a target fragment is inserted into a specified site in the genome, or a specified fragment in the genome is deleted or replaced, thereby achieving high targeting accuracy.

Description

[Technical Field]

[0001] The present invention relates to the field of biotechnology, and to gene editing technology, particularly to gene editing technology mediated by DNA, RNA or RNP pathways and its applications. [Background technology]

[0002] Currently, the main gene editing technologies in the biotechnology field are ZFN, TALEN, CRISPR / Cas9, and Targetron technologies. Among them, ZFN technology was the first to be developed. However, its DNA-binding domain can only recognize sequences up to 9 bp in length, greatly limiting its targeting accuracy in practical applications. Furthermore, the actual design of this technique is tedious, and it cannot knockout sequences with unknown upstream and downstream sequences. Furthermore, its cytotoxicity and off-target rate are high. Compared to ZFN, TALEN is simpler to design and can recognize 17-18 bp sequences with higher specificity. However, because the core technology is controlled by individual commercial companies, most laboratories are unable to complete it independently, limiting its widespread use and application to some extent. At the same time, its construction process is still relatively complex. CRISPR / Cas9 is the simplest and most easily operable of the three. First discovered in archaea and bacteria, it specifically recognizes a sequence of approximately 20 bp, causing a double-strand break at a specific site through the action of the Cas9 endonuclease, which then repairs the break via the system's own DNA repair function, allowing for gene editing operations. However, all three techniques suffer from off-target issues, even the CRISPR / Cas9 technique, which has the longest recognition length. Research has shown that the specificity of CRISPR / Cas9 recognition of target sites primarily depends on pairing within 10–12 base pairs near the gRNA PAM site, which tends to cause nonspecific cleavage. Furthermore, after generating DNA double-strand breaks using these three techniques, the DNA double-strand breaks are likely repaired by non-homologous end joining rather than homologous recombination, resulting in the generation of random sequences. This uncertainty significantly impairs their practical application, especially in humans and clinical settings. At the same time, these three techniques introduce genetic material and proteins that are not part of the recipient system, such as humans, which can also have unexpected effects. Meanwhile, the updated Targetron technology uses group II introns to insert sequences into specific sites in the genome and mutate the corresponding gene. However, this technology inevitably causes double-strand breaks in the genome, introducing exogenous group II introns into the genome to create a "scar." Furthermore, because this technology is derived from prokaryotes, the RNA produced for reverse transcription lacks transmembrane transport function, limiting its application. Furthermore, and most importantly, while this technology has worked well in the field of bacterial gene editing, it has not worked well in higher organisms. All four gene editing technologies require the introduction of proteins or nucleic acids that do not belong to the receptor system, increasing uncertainty about their effectiveness and significantly hindering their clinical application. Summary of the Invention

[0003] In order to solve the above problems, the object of the present invention is to provide a gene transcription framework, which, after transcription, can be transported to the cell nucleus or cytoplasm via DNA, RNA and / or RNP pathways to insert a target fragment into a specific site in the genome, or delete or replace a specific fragment in the genome, and at the same time has higher target accuracy. Another object of the present invention is to provide a vector system that can be mediated by the DNA, RNA and / or RNP pathways. A third object of the present invention is to provide a gene editing method that uses DNA, RNP, or RNA (which can be produced in vitro) and related proteins as a medium to be transferred to the cell nucleus or cytoplasm via the DNA, RNA, or RNP pathway, and inserts a target fragment into a specific site in the genome, or deletes or replaces a specific fragment in the genome, while at the same time achieving higher target accuracy, on the premise that the gene editing method minimizes the introduction of foreign systems or substances and does not cause double-strand breaks. In order to achieve the above object, the present invention provides a gene transcription framework, the gene transcription framework including, in a 5'→3' direction, a target site upstream sequence, a sequence to be inserted, and a target site downstream sequence; A gene transcription framework characterized in that the gene transcription framework is a DNA sequence that can be transcribed by RNA polymerase I, RNA polymerase II, or RNA polymerase III, the target site upstream sequence or its complementary sequence in a transcription product of the gene transcription framework or a transformation product of the transcription product is capable of hybridizing to the upstream sequence of a corresponding target site in a cell genome or its complementary sequence, the target site downstream sequence or its complementary sequence is capable of hybridizing to the downstream sequence of a corresponding target site in the cell genome or its complementary sequence, the target site upstream sequence and the target site downstream sequence are directly connected in the corresponding target gene sequence in the genome, and the site between the target site upstream sequence and the target site downstream sequence in the target gene sequence on the genome is the target site for the sequence to be inserted. The gene transcription framework provides a gene transcription framework that is used to insert an insert sequence into a genomic target site. Preferably, the cell is a eukaryotic cell. The present invention further provides a vector system, the vector system comprising one or more types of vectors, the one or more types of vectors comprising: 1) one or more gene transcription frameworks according to claim 1; 2) one or more short interspersed element sequences and / or partial short interspersed element sequences and / or similar short interspersed element sequences; and / or one or more 3) long interspersed element sequences and / or ORF1p coding sequences and / or ORF2p coding sequences; Including, wherein the components 1), 2) and / or 3) are located in the same or different vectors of the vector system; If there are multiple components 1), they may be located in the same or different vectors of the vector system; If there are multiple components 2), they may be located in the same or different vectors of the vector system; If there are multiple components 3), they may be located in the same or different vectors of the vector system; the vector comprises one or more promoters, the promoters being RNA polymerase I promoter, RNA polymerase II promoter, or RNA polymerase III promoter, and located upstream of the components 1), 2), and / or 3); The vector system is provided, wherein the vector system is mediated by a DNA, RNA and / or RNP pathway. Furthermore, the vector is a eukaryotic expression vector, a prokaryotic expression vector, a viral vector, a plasmid vector, an artificial chromosome, a phage vector, or a cosmid vector. Furthermore, the vector is an expression vector, a cloning vector, a sequencing vector, a transformation vector, a shuttle vector or a multifunctional vector. The vector system is further characterized in that, when 1) and 2) are located in the same vector, the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence are located downstream of the gene transcription framework, and the gene transcription framework is directly or indirectly connected to the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence, and when directly connected, the gene transcription framework shares a promoter with the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence, and when indirectly connected, the gene transcription framework shares a promoter or does not share a promoter with the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence. The vector system is further characterized in that, when 1) and 3) are located in the same vector, the one or more long interspersed element sequences, and / or one or more ORF1p coding sequences, and / or one or more ORF2p coding sequences are located upstream and / or downstream of the gene transcription framework, and the gene transcription framework is directly or indirectly connected to the one or more long interspersed element sequences, and / or one or more ORF1p coding sequences, and / or one or more ORF2p coding sequences; when directly connected, the gene transcription framework shares a promoter with the one or more long interspersed element sequences, and / or one or more ORF1p coding sequences, and / or one or more ORF2p coding sequences; when indirectly connected, the gene transcription framework shares a promoter or does not share a promoter with the one or more long interspersed element sequences, and / or one or more ORF1p coding sequences, and / or one or more ORF2p coding sequences. Furthermore, when 1), 2) and 3) are located in the same vector, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are located downstream of the gene transcription framework and / or downstream of the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, and when the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are located downstream of the gene transcription framework, the long interspersed element sequence and / or ORF1p coding sequence are located downstream of the gene transcription framework. sequence and / or ORF2p coding sequence is located upstream of the gene transcription framework, and / or the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence is located downstream of the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence, and the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence is located downstream of the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, a vector system characterized in that the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence are located downstream of the gene transcription framework, and the gene transcription framework, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are directly or indirectly connected to the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence; when directly connected, the gene transcription framework, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence share a promoter with the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence; when indirectly connected, the gene transcription framework, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence share a promoter or do not share a promoter with the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence. Furthermore, when 2) and 3) are located in the same vector, the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence are located upstream and / or downstream of the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence, and the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are directly or indirectly connected to the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, or directly connected to the above. A vector system characterized in that when the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence share a promoter with and are indirectly connected to the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence may or may not share a promoter with the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence. To improve transcription efficiency, the number of gene transcription frameworks, short interspersed element sequences, partial short interspersed element sequences, and / or similar short interspersed element sequences, long interspersed element sequences, and / or ORF1p coding sequences and / or ORF2p coding sequences in the vector system is increased. Although short interspersed element sequences and long interspersed element sequences naturally exist in different species, the activity of the native promoters is low. Therefore, transcription by additional promoters and / or additional transcription of short interspersed element sequences, partial short interspersed element sequences, and / or similar short interspersed element sequences, and / or long interspersed element sequences and / or ORF1p coding sequences and / or ORF2p coding sequences is applied to improve expression and gene editing efficiency. The present invention further provides a method for editing a genome sequence, comprising: 1) A step of selecting a site (target site) into which a target gene to be edited is inserted from the genome, and determining the upstream sequence (target site upstream sequence, target site upstream sequence) and downstream sequence (target site downstream sequence, target site downstream sequence) of the insertion site of the target gene on both sides of the insertion site; 2) a process for producing the vector system; 3) transforming or transfecting a cell, tissue, organ, embryo, or organism with the vector system to achieve gene editing; The present invention provides a method for editing a genome sequence, comprising: The present invention further provides the use of the above vector system in inserting, deleting, or substituting DNA sequences in any region of the genome. Preferably, the DNA sequence is one or more of a CNV sequence, a CNV terminal sequence, a short interspersed element sequence, a partial short interspersed element sequence, a similar short interspersed element sequence, a long interspersed element sequence, a partial long interspersed element sequence, an ORF1p coding sequence and / or an ORF2p coding sequence. If the DNA sequence is a CNV sequence or a CNV end (end of CNV, CNV end) sequence, the CNV end (i.e., between the gene portion at the CNV end and the partial SINE (partial SINE portion, partial SINE sequence)) can be edited to insert a sequence that is not homologous to the genome or local genome to block copy number changes and expression changes of the gene, or the gene partial sequence at the CNV end can be deleted to change the corresponding cellular expression. The CNV end includes the gene portion and the partial SINE. The present invention further provides use of the above vector system as a pharmaceutical for the prevention and / or treatment of cancer, gene-related genetic diseases, and neurodegenerative diseases. Preferably, the cancer is glioma, breast cancer, cervical cancer, lung cancer, gastric cancer, colon cancer, duodenal cancer, leukemia, prostate cancer, uterine cancer, thyroid cancer, lymphoma, pancreatic cancer, liver cancer, melanoma, skin cancer, pituitary tumor, germ cell tumor, meningioma, meningeal carcinoma, glioblastoma, various astrocytomas, various oligodendrogliomas, astrooligodendrogliomas, various ependymomas, choroid plexus papilloma, choroid plexus carcinoma, chordoma, neuroblastoma, various ganglioneuromas, olfactory neuroblastoma, sympathetic neuroblastoma, pineocytoma, pineoblastoma, medulloblastoma, trigeminal neurinoma, acoustic neuroma, jugular vein tumor, jugular globus tumor, hemangioplexioma, craniopharyngioma, granulosa cell tumor, and / or granulosa cell tumor. Preferably, the gene-related genetic disease is Huntington's disease, fragile X syndrome, phenylketonuria, pseudohypertrophic progressive muscular dystrophy, mitochondrial encephalomyopathies, spinal muscular atrophy, parkinsonian superposition syndrome, parkinsonism, Parkinson's disease, albinism, red-green color blindness, red-green color dysphoria, achondroplasia, alkaptonuria, congenital hearing loss, thalassemia, sickle cell anemia, hemophilia, epilepsy, myoclonus, dystonia, stroke and schizophrenia associated with genetic changes, vitamin D-resistant rickets, familial colonic polyposis, hereditary nephritis. Preferably, the neurodegenerative disease is Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar ataxia, multiple system atrophy, primary lateral sclerosis, Pick's disease, frontotemporal dementia, dementia with Lewy bodies, or progressive supranuclear palsy. The present invention can prevent the occurrence of the above cancers and their metastases, inhibit their growth, prevent their progression, or reverse their nature to cure them. It can also prevent, delay, or improve drug resistance to insulin, levodopa, various tumor chemotherapeutic drugs, and targeted drugs, and slow or stop genetic and state changes, tissue and organ regeneration, and biological regeneration in cells, tissues, organs, embryos, or organisms. By applying the technology related to the present invention, copy number variations (CNVs) and their terminal portions (CNV terminals) in each gene can be edited to change the terminal position or stabilize the terminal. Because copy number variations in each gene and its terminal portion (CNV terminal) determine gene expression, changing the terminal position or stabilizing the terminal can stabilize or change the state of cells, tissues, embryos, organs, and organisms. This can be used in technical fields that modify or change the genes and states of cells, tissues, organs, embryos, and organisms. This can be used to modify the genomes of organisms such as humans to improve function, to modify the genomes of organisms such as humans to treat various gene-related genetic diseases such as Huntington's disease and Fragile X syndrome, to slow or stop changes in genes and states of cells, tissues, organs, embryos, and organisms (such as aging), and to change the genes and states of cells, tissues, organs, embryos, and organisms. Reconstruction and biological regeneration of embryos, tissues or organs, assisting reproduction by introducing transcription factors that transform somatic cells into germ cells, preventing or delaying neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple system atrophy, primary lateral sclerosis, spinocerebellar ataxia, Pick's disease, frontotemporal dementia, Lewis body dementia, and progressive supranuclear palsy, inhibiting the metabolic activity and proliferation rate and production of tumor cells, slowing their progression and improving their malignancy, research and treatment of all other diseases associated with genetic and CNV changes such as diabetes, and other physiological, pathological, and pathophysiological studies. In the present invention, the sequence inserted into the gene transcription framework can be an exogenous sequence or an endogenous sequence, and the length of the one-time insertion sequence is 1 bp to 2000 bp. Insertion of a DNA sequence of any length into the genome can be achieved by performing multiple insertions. The nucleotide sequence length of the target site upstream sequence (the sequence upstream of the target site) may be between 10 bp and 2000 bp, and the nucleotide sequence length of the target site downstream sequence (the sequence downstream of the target site) may be between 10 bp and 2000 bp. Features of the present invention: This invention allows the insertion sequence to be inserted into a specific site in the genome by relying on the upstream and downstream sequences of the target site on both sides of the insertion sequence on the vector, and then positioning it at the insertion site on the genome. The insertion sequence is inserted into the selected site in the genome with the help of short interspersed elements, long interspersed elements, and their expression proteins. Furthermore, ORF2p expressed in cells or by a vector can smoothly slide from the 3' end of the nucleic acid vector to the single-strand break site in the genome only if the upstream sequence of the target site on the vector or its complementary sequence perfectly matches the upstream sequence of the target site on the cellular genome or its complementary sequence. This significantly improves targeting accuracy, avoids unexpected breaks, and theoretically achieves higher targeting accuracy than existing gene editing technologies. Furthermore, the necessary RNA and corresponding endogenous proteins, such as ORF1p and ORF2p, can be generated in vitro, allowing target sequences and genes in the genome to be edited via the RNA or RNP pathway, transfecting them directly into the nucleus without the need for DNA fragments. The RNA or RNP pathway minimizes the impact on receptor systems, reducing nonspecific effects and improving targeting accuracy. The nuclear localization function of ORF1p and ORF2p, along with ORF1p's protective effect on nucleic acids, allows transfected RNA and proteins to be guided to the nucleus, making this useful for gene editing in cells that are difficult to manipulate due to the difficulty of vector transport into the nucleus. The present invention has better targeting and does not generate double-strand breaks, allowing for more accurate target sequence identification and cleavage, and allows for the directionally inserting of desired sequences through homologous recombination, and the deletion and replacement of corresponding fragments.However, existing gene editing technologies such as CRISPR cause double-strand breaks and unpredictable random mutations, making it unlikely that target sequences can be introduced through extremely inefficient homologous recombination.The present invention does not generate double-strand breaks, eliminating the risk of DNA double-strand breaks and the introduction of unexpected random sequences. The present invention allows for the incremental introduction of long sequences into a genome by continually designing vectors for further insertion according to new sites generated after previous insertions, which is also difficult to achieve with currently known gene editing technologies. Similarly, targeted, precise sequence deletion and sequence replacement in the genome, which is difficult to achieve with conventional technologies, can also be achieved with the present invention. Furthermore, manipulations such as editing or stabilizing CNVs or their ends to change or stabilize gene expression and / or state of cells or organisms, which cannot be achieved with existing gene editing technologies, can be achieved with the present invention. The beneficial effects of the present invention are as follows: The present invention provides a gene editing method based on the eukaryotic retrotransposition mechanism, which can be mediated by DNA, RNA, or RNP pathways. This method utilizes the inherent mechanisms of eukaryotes, minimizes the introduction of exogenous systems or substances, and does not generate double-strand breaks. This method is mediated by DNA, RNP, or RNA (which can be prepared and produced in vitro) and associated proteins (which can be prepared and produced in vitro). By introducing the desired fragment (the inserted sequence) into the nucleus or cytoplasm via the DNA, RNA, or RNP pathway, the desired fragment (the inserted sequence) can be inserted into a selected locus within the genome, or the selected fragment within the genome can be deleted or replaced, while still achieving high targeting accuracy. Because this method does not require the introduction of exogenous systems such as prokaryotic proteins and does not generate double-strand breaks, it is easier to apply clinically than existing gene editing technologies. [Brief explanation of the drawings]

[0004] [Figure 1] This is a basic schematic diagram of gene editing based on the retrotransposition mechanism possessed by eukaryotes themselves. [Figure 2] 1 shows a schematic diagram of DNA-mediated genomic insertion or deletion. [Figure 3] 1 shows a schematic diagram of RNA- and RNP-mediated genomic insertion and deletion. [Figure 4]1 shows a schematic diagram of blocking alterations at the CNV ends by inserting non-homologous sequences at the CNV ends. [Figure 5] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention. [Figure 6] 1 is a schematic diagram of the structure of a gene transcription framework-linked promoter provided by the present invention. [Figure 7] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention, which links a promoter and a short interspersed element, a partial short interspersed element, or a similar short interspersed element. [Figure 8] 1 is a schematic diagram of the structure of a promoter linked upstream of the gene transcription framework provided by the present invention, and a long interspersed element or ORF1p coding sequence or ORF2p coding sequence linked downstream of the gene transcription framework provided by the present invention. [Figure 9] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention, in which the gene transcription framework is linked upstream with a long interspersed element or ORF1p coding sequence or ORF2p coding sequence, and a promoter is linked upstream of the long interspersed element or ORF1p coding sequence or ORF2p coding sequence. [Figure 10] A schematic diagram of the structure of the gene transcription framework provided by the present invention, in which the promoter is linked upstream of the gene transcription framework, a short interspersed element or partial short interspersed element or similar short interspersed element is linked downstream of the gene transcription framework, and a long interspersed element or ORF1p coding sequence or ORF2p coding sequence is connected downstream of the short interspersed element or partial short interspersed element or similar short interspersed element. [Figure 11] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention, in which the gene transcription framework is connected to a short interspersed element, a partial short interspersed element, or a similar short interspersed element downstream, the gene transcription framework is linked to a long interspersed element or an ORF1p coding sequence or an ORF2p coding sequence upstream, and the upstream of the long interspersed element or the ORF1p coding sequence or the ORF2p coding sequence is linked to a promoter. [Figure 12] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention, in which the gene transcription framework and the short interspersed elements, partial short interspersed elements, or similar short interspersed elements do not share a promoter. [Figure 13] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention, in which the gene transcription framework and the long interspersed element or the ORF1p coding sequence or the ORF2p coding sequence do not share a promoter. [Figure 14] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention. The gene transcription framework, short interspersed elements, partial short interspersed elements or similar short interspersed elements, and long interspersed elements, ORF1p coding sequence or ORF2p coding sequence do not share a promoter, while the short interspersed elements, partial short interspersed elements or short interspersed-like elements, and long interspersed elements or ORF1p coding sequence or ORF2p coding sequence share a promoter. [Figure 15] 1 is a schematic diagram of the structure of the gene transcription framework provided by the present invention. The gene transcription framework, short interspersed elements, partial short interspersed elements or similar short interspersed elements, and long interspersed elements, ORF1p coding sequence or ORF2p coding sequence do not share a promoter, while the short interspersed elements, partial short interspersed elements or short interspersed-like elements, and long interspersed elements or ORF1p coding sequence or ORF2p coding sequence share a promoter. [Figure 16] 1 shows a plasmid map of the plasmid pSIL-eGFP-VEGFA1-Alu1 constructed in Example 1, in which the gene transcription framework VEGFA1 is inserted into the vector of Example 1. [Figure 17] 1 shows a plasmid map of the plasmid pBS-L1PA1-CH-mneo-IT15-1 constructed in Example 6, in which the gene transcription framework IT15-1 was inserted into the vector in Example 6. Modes for carrying out the expression

[0005] This study is based on a transposon-mediated genome reorganization mechanism ubiquitous in eukaryotes, which alters gene copy number and repeat sequences. This mechanism can result in the deletion or addition of pathogenic trinucleotide repeats in several central nervous system degenerative diseases, such as Huntington's disease and Fragile X syndrome. This mechanism coincides with homologous recombination, which can be inhibited by methylation and correlates with expression levels. (Figure 1) In FIG. 1, the present invention relates to short interspersed elements (SINEs), long interspersed elements (LINEs) and their associated proteins, such as open reading frame 1 protein (ORF1p), open reading frame 2 protein (ORF2p) and other types of open reading frame proteins (ORFp). Sine-interspersed elements (SINEs) mainly include Alu and SVA elements in primates, various mammalian-wide interspersed repeat elements (MIRs) such as MIR and MIR3, Mon-1 in monothrenosum, B1 and B2 elements in rodents, the HE1 family in zebrafish, Anolis SINE2 and Sauria SINE in reptiles, IdioSINE1, IdioSINE2 in invertebrates such as squid, Sepiasine, Sepios-SINE1, Sepios-SINE2A, Sepios-SINE2B and Egopsine, and p-SINE1 in plants such as rice.Long-interspersed elements (SINEs) primarily include various LINE-1 (L1), various LINE-2 (L2), and various LINE-3 (L3), including R2, RandI, L1, RTE, I, and JOCV elements in various species of organisms, as well as other LINE species. These structures are widely found in various plants and animals and are scattered throughout the genome, each with its own specific SINEs and LINEs corresponding to its complementary functions. SINEs are primarily characterized by relatively short transposons distributed throughout the genome, containing internal RNA polymerase III promoters and ending in A- or T-rich tails or short simple repeats. They are reverse-transcribed with the aid of LINEs, and the right half of their transcripts contains a functional structure capable of inversion. LINEs, on the other hand, are characterized by transposons widely distributed throughout the genome that contain reverse transcriptase coding sequences. Both SINEs and their corresponding LINEs continuously remodel the genome through similar mechanisms. The basic principle of this mechanism is that the lariat structure generated in vivo by pre-mRNA processing is connected to the right half of the residual reverse-transcribed structure of the SINE by shearing. (The right half of a SINE with a reverse-transcribed structure generated by a complete SINE transcript cleaved at the midpoint is called a partial SINE sequence, and the shearing site varies depending on the SINE species.) The natural shearing site of a SINE is generally located in the middle of its entire length. For SINEs with a typical total length of approximately 100-400 nt, the natural shearing site is usually located at 100-250 nt. For example, for an Alu element with a total length of approximately 300 bp, the shearing site is located at 118 nt. For various types of MIRs with a total length of approximately 260 nt, the shearing site can be observed in the range of 100-150 nt. In fact, regardless of the location of the sheared site, as long as it is sheared, the remaining right-hand portion contains a complete reverse transcription functional structure (its secondary structure forms a special structure, usually Ω-shaped; its primary structure is characterized by a sequence containing two segments separated by an intermediate spacer sequence between them, which can bind complementary sequences in the genome without the intermediate spacer sequence and directly connect the two sequences. ORF2p, encoded by the LINE, binds to the sequence 3' of the two sequences in the transcript, cleaves the genomic single strand at the corresponding genomic locus between the two sequences, and initiates reverse transcription), i.e., a partial SINE sequence. Certain SINEs, such as partial SINEs generated by Alu elements, are recorded as partial Alu, specifically its intermediate adenylate repeat, or two to three bases upstream, and an Alu right monomer and a 3' poly(A) repeat following the right monomer. Furthermore, sequences that contain reverse transcription functional structures and can initiate reverse transcription but differ in sequence from conventional SINEs are called similar short interspersed elements (similar SINEs). Proteins (e.g., ORF1p and ORF2p) expressed in corresponding types of LINEs (e.g., LINE-1, which corresponds to the function of Alu elements, and LINE-2, which corresponds to various MIR elements) convert RNA into double-stranded DNA and bind to its complementary sequence in the genome (the transcribed RNA is reverse transcribed to generate single-stranded DNA, and the double-stranded DNA generated by using the genomic sequence as a primer is the "transcription product"). By forming a specific Ω structure, exogenous sequences are inserted into the genome by homologous recombination. Furthermore, LINEs can also transcribe downstream sequences (i.e., 3' transduction) and bind to complementary sequences in the genome to form Ω structures, thereby converting the RNAs described above into double-stranded DNA and inserting them into the genome. Taking Alu elements and their functionally complementary counterparts, LINE-1, as an example, they can cleave pre-mRNAs generated after gene expression to generate lariats with overlapping sequences that can occur anywhere in the pre-mRNA. The difference lies in the strength of the cuts used to generate these lariats. The cleavage strength (based on sequence differences) of lariats upstream and downstream of an exon is higher than that of other surrounding lariats, allowing them to easily and completely cleave the exon during the pre-mRNA process, inhibiting the production of other lariats. At the same time, ORF1p produced by LINE-1 can protect the nucleic acid bound to it and, together with ORF2p produced by LINE (LINE-1), place the bound nucleic acid in the nucleus and transport it into the nucleus. Furthermore, ORF2p can bind to a special Ω secondary structure in Alu element transcripts and mediate subsequent genomic single-strand cleavage, reverse transcription, and integration into the genome. Alu element transcripts can be cleaved at a specific site (i.e., the small cytoplasmic element RNA produced by multiple replacement loci) (i.e., the following scAlu cleavage site or the natural cleavage site, which is generally located before the middle poly(A) sequence of the Alu transcript, but this may vary depending on the actual situation), generating a small cytoplasmic Alu (scAlu) RNA and the remaining portion containing the right monomer (reverse-transcribed functional ORF2p). The remaining portion containing the right monomer is referred to as partial Alu RNA, and the DNA sequence from which the partial Alu RNA is transcribed is referred to as partial Alu sequence (partial Alu). The resulting lariat structure can then be ligated from its 3' end to the remainder of the truncated Alu transcript, including the reverse transcription functional structure. ORF2p is recruited via an adenylate-rich sequence and can bind to the 3' leg of the Ω bipod formed by the partial Alu RNA secondary structure. A sequence in the genome that matches the sequence in the Ω bipod (primarily the UU / AAAA discontinuity between U and A in the partial Alu RNA, i.e., the gap) is identified. A single strand of the genome opposite the Ω gap is cleaved, and a genomic sequence complementary to the Ω bipod sequence is melted and used as a primer for reverse transcription. This process is called target-primed reverse transcription (TPRT). ORF2p then moves to the 3' end of the single-stranded DNA generated by reverse transcription, where it binds to a complementary sequence on the genome and forms an Ω secondary structure at the corresponding insertion site (target site) on the genome (since the insertion site (target site) does not contain the sequence to be inserted, whereas the sequences on both sides of the sequence to be inserted into the single-stranded DNA exist on both sides of the insertion site (target site) on the genome). ORF2p slides along the complementary sequence from 3' to 5' into the Ω structure, identifying a genome sequence with six nucleotides (mainly the four nucleotides at the 3' and two nucleotides at the 5') that are complementary to the bottom two legs of the Ω structure, generating double-stranded DNA through a similar process. Note that only perfectly matched sequences can slide ORF2p into the incision site (i.e., the site on the genome corresponding to the gap in the Ω structure on the generated DNA), ensuring its targeting accuracy. The final double-stranded DNA then reconnects to both sides of the corresponding insertion site (target site) and forms an Ω structure again. If six nucleotides (mainly between the four nucleotides at the 3' and two nucleotides at the 5' recognized by ORF2p) are discontinuous in the gap (the Ω gap in the double-stranded DNA structure), the endonuclease action of ORF2p can create two single-stranded DNA incisions in the gene (genome) corresponding to the gap and in the other strand of the generated double-stranded DNA. The central circular portion of the Ω structure is then inserted into the genome by homologous recombination. By modifying the inserted sequence, other effects such as deletions or substitutions can be achieved in the genome through homologous recombination. In the above process, the annealing and deconstruction functions of ORF1p, encoded by the LINE, stabilize the secondary structures generated by the nucleic acid in the described genome reorganization process and its binding to the genome, thereby promoting its separation from the genome after binding and action. ORF1p also possesses high RNA affinity and nuclear localization functions. ORF2p can only cleave one strand of the genomic duplex and does not generate double-strand breaks, making it highly safe. A similar mechanism applies to other SINE-LINE combinations. Changes in local copy number variation during physiological and pathological processes such as embryonic development and tumorigenesis, as well as the preference for SINE sequences inserted into the human genome, are manifestations of this mechanism in nature. It has been reported that transcribed mRNA sequences can be integrated into the genome with the aid of ORF1p and ORF2p. However, because the transcription template was a purely exogenous, non-homologous sequence, it was unable to target specific sites within the genome and fragments with reverse transcription functional structures were not attached, resulting in an inefficient, random process that was difficult to control. According to the present invention, the transcription sequence is redesigned and linked to sequences containing reverse transcription functional structures, such as various types of SINEs or partial SINEs, by various active or passive means, achieving more precise and efficient gene editing effects. Under physiological conditions, copy number variations (CNVs) resemble copies of intact genes. Through the above mechanisms, CNVs can be continuously extended like copies of intact genes, constantly changing protein expression and various conditions in cells, tissues, and organisms. CNV termini consist of an upstream gene segment and a downstream partial SINE sequence segment. Short sequence fragments contained in lariats are continuously inserted between the two CNV termini via the lariat sequence linked to the partial SINE sequence, extending the CNV. During early embryonic development, LINE transcription significantly increases, and genomic SINEs (e.g., Alu element sequences) show significant demethylation. LINE-mediated 3' transduction (based on the deletion of the right monomer of a SINE upstream of the promoter of the corresponding (related) gene and the complete SINE structure downstream) initiates the extension of the copy number variation (CNV) of the associated gene, but homologous recombination of the demethylated SINE sequence deletes most of the previously extended CNV (initiation). Subsequently, fully reprogrammed embryonic cells regain a hypermethylated state, and the partial SINE sequences at the CNV ends mediate the gradual extension of the CNV ends, thereby changing the expression status and state of each cell. The gene expression status of each cell then influences the CNV changes via the lariat, thus leading to genomic changes and stepwise differentiation induction. This is consistent with the general changes in CNVs in embryos and the differences in CNVs in various tissues. Long CNVs in various genes (e.g., CNVs in oncogenes) are commonly present in all types of tumor cells and are positively correlated with clinical grade. At the same time, the expression levels of proto-oncogenes and tumor suppressor genes are also directly proportional to the length of the CNVs. Therefore, tumor formation and progression should be related to CNV abnormalities in proto-oncogenes or tumor suppressor genes. Furthermore, some irreversible diseases associated with external stimuli, such as diabetes, are also associated with CNV abnormalities. Since the majority of drug resistance is associated with changes in the expression of corresponding proteins due to long-term external stimuli, CNV changes in those corresponding genes may be involved, and this technology can be used to improve or prevent drug resistance. I. DNA-mediated genome sequence insertion editing technology (If gene editing is mediated by DNA, 1–40 TTAAAA or TTTTAA sequences can be added to the plasmid to assist in the conversion of RNA to DNA) (as shown in Figure 2) 1. Lasso-based approach: Select sequences (within 2000 bp) upstream and downstream of the insertion site (i.e., the target site), and add the insertion sequence (within 2000 bp) to the intermediate insertion site between the upstream and downstream sequences. The designed sequence is synthesized, incorporated into the vector, and transcribed by RNA polymerase II. Other regions of the vector can be inserted using "SINE sequences" (0 to 20; to reduce the impact on the receiving system, SINE sequences can be used for the corresponding species of the receiving system). SINE sequences can be Alu sequences in primates or Mon-1 sequences in monotremes, and the number of SINE sequences within a certain range is proportional to the efficiency. It is also possible to use non-naturally occurring, non-species SINEs, after which a LINE or its protein-coding sequence must be introduced that corresponds to the function of the SINE used (see below) and is initiated by RNA polymerase II or III. A corresponding termination signal for RNA polymerase is optionally linked after the SINE sequence. (A corresponding LINE sequence or a protein sequence encoded by the LINE sequence can be optionally inserted before the termination signal after the SINE sequence, such as a LINE-1 sequence corresponding to an Alu element in the SINE, or the ORF1p and ORF2p sequences encoded by the LINE-1 sequence.) A protein required for gene editing can be expressed, such as a LINE-2 sequence corresponding to an MIR element, or the ORF1p and ORF2p sequences encoded by the LINE-2 sequence, to achieve gene editing or improve editing efficiency. (When transcription is initiated by RNA polymerase II, the corresponding termination signal is a polyA sequence, and its length can be appropriately extended (within 200 bp) to increase the recruitment of ORF2p. In other cases, a polyA sequence of an appropriate length can be added (within 200 bp) after the ORF1p and ORF2p sequences preceding the termination signal, and the polyA sequence at the end of the SINE sequence can be appropriately extended (within 200 bp).) Then, the vector (other vectors expressing the LINE sequence corresponding to the SINE sequence contained in the vector or the protein sequence encoded by the LINE sequence, for example, the LINE-1 or ORF2p and / or ORF1p sequence corresponding to the Alu sequence) can be simultaneously transfected into the recipient system to improve gene editing efficiency. If the recipient system does not express the above-mentioned proteins, such as ORF1P or ORF2p of the corresponding LINE, additional protein expression is required. In addition, a vector expressing a SINE can also be simultaneously transfected into the recipient system to improve gene editing efficiency) is transferred to cells and tissues cultured in vitro via conventional means such as liposome or viral transfection, or administered to an organism via routes such as blood, lymph, cerebrospinal fluid, or local tissue, and the constructed vector enters the nucleus for expression and completes the purpose of inserting the corresponding sequence to be inserted into the corresponding site on the genome. (If the expected efficiency is not achieved, it may be due to inefficient formation of a lasso structure consisting of the upstream and downstream sequences of the insertion site (target site) and the intermediate sequence to be inserted.) The length of the upstream and downstream sequences or the inserted sequence can be increased or decreased to facilitate the formation of a lariat structure. Alternatively, a lariat structure containing the insertion site (target site) can be detected using the following detection method, and the sequence or partial sequence of the lariat structure can be used as the upstream and downstream sequences of the target site. The upstream and downstream sequences of the target site can then be appropriately extended according to the genome sequence. The inserted sequence can be placed between the upstream and downstream sequences of the target site to form a gene transcription framework, which can be constructed into a vector. Transfection of this vector can also improve gene editing efficiency. (The constructed vector can be incubated in a physiological solution containing ORF1p and / or ORF2p for a short period of time (at an appropriate temperature, such as room temperature or 37°C, for up to 48 hours) to improve the nuclear localization and import efficiency of the vector.)By continuing genome insertion according to the above method based on the new site generated after insertion, sequence insertion can be performed continuously, and long fragment insertion into the genome can be achieved without significant length restrictions. 2. SINE (sequence) direct connection approach: This approach does not require the generated lasso to be connected to the cleaved SINE transcript. During vector construction, the upstream and downstream sequences of the insertion site (target site) and the sequence to be inserted between them are directly connected to the SINE-related sequence. This method is suitable for prokaryotes such as bacteria, which cannot generate lasso structures without the eukaryotic pre-mRNA splicing machinery. It is also suitable for eukaryotes that possess the pre-mRNA splicing machinery. This also applies to the LINE-mediated approach described below. The specific steps are as follows: The upstream and downstream sequences (within 2000 bp) of the insertion site (target site) and the sequence to be inserted between the upstream and downstream sequences (within 2000 bp) form the gene transcription framework, which is initiated by a promoter for RNA polymerase II or III, followed by synthesis of a sequence connecting the SINE sequence, partial SINE sequence, or similar SINE sequence. (Optionally, to improve gene editing efficiency, a LINE sequence or its protein-encoding sequence that can support the corresponding SINE function can be added after the SINE sequence, partial SINE sequence, or similar SINE sequence. If the recipient system does not express the LINE or its encoded protein itself, the encoded protein must be added to the recipient system or additionally expressed in the recipient system.) Then, a termination signal for the corresponding type of RNA polymerase can be selectively added after the SINE sequence, partial SINE sequence, or similar SINE sequence (if the termination signal is a polyA sequence, the termination signal can be appropriately extended (within 200 bp) to increase the recruitment of ORF2p). Alternatively, to increase the recruitment of ORF2p, a polyA sequence of an appropriate length (within 200 bp) can be added after the SINE sequence, partial SINE sequence, similar SINE sequence, LINE sequence, ORF1p sequence, and / or ORF2p sequence and before the termination signal, and the sequence can be constructed into a vector. The vector is transferred to cells and tissues cultured in vitro via conventional means such as liposome or viral transfection, or administered to an organism via routes such as blood, lymph, cerebrospinal fluid, or local tissue. (The constructed vector can be incubated in a physiological fluid containing ORF1p and / or ORF2p for a short period of time (at an appropriate temperature, such as room temperature or 37°C, for up to 48 hours) to improve the nuclear localization and import efficiency of the vector.) The constructed vector enters the nucleus for expression and completes the insertion of the corresponding sequence into the corresponding site on the genome. By continuing genome insertion according to the above method based on the new site generated after insertion, sequence insertion can be performed continuously, and long fragment insertion into the genome can be achieved without significant length restrictions. LINE mediated approach: RNA polymerase II initiates expression of the LINE or its protein ORF2p and / or ORF1p coding sequences, followed by a sequence designed in the same manner as the SINE sequence direct splicing approach. (To minimize impact on the gene editing recipient system, the SINE and LINE types of the recipient system can be selected. To improve efficiency, the SINE sequence within the LINE can be selected to functionally correspond to the previous LINE.) The RNA polymerase II termination signal used can then be selectively ligated to the end. The vector is transferred to in vitro cultured cells and tissues via conventional means, such as liposome or viral transfection, or administered to living organisms via routes such as blood, lymph, cerebrospinal fluid, or local tissue. (The constructed vector can be incubated in physiological fluids containing ORF1p and / or ORF2p for a short period of time (at an appropriate temperature, such as room temperature or 37°C, for up to 48 hours) to improve vector nuclear localization and import efficiency.) The constructed vector enters the nucleus for expression and completes its purpose of inserting the corresponding sequences into the corresponding genome insertion sites. By continuing genome insertion according to the above method based on the new site generated after insertion, sequence insertion can be performed continuously, and long fragment insertion into the genome can be achieved without significant length restrictions. Downstream binding ORF2p binding sequence approach: The Ω structure formed by the binding of the upstream sequence of the target site, downstream sequence, and intermediate insertion sequence of the gene transcription framework on the vector to the genome replaces the SINE reverse transcription functional structure and initiates reverse transcription. An ORF2p binding sequence (e.g., a poly(A) sequence) is attached downstream of the downstream sequence of the target site in the gene transcription framework. The LINE sequence, ORF1p, and / or ORF2p coding sequences can be optionally added to the same vector as the gene transcription framework or to a separate vector to improve efficiency. The vector is transferred to cells and tissues cultured in vitro via conventional means such as liposome or viral transfection, or administered to an organism via routes such as blood, lymph, cerebrospinal fluid, or local tissue. (The constructed vector can be incubated in a physiological fluid containing ORF1p and / or ORF2p for a short period of time (at an appropriate temperature, such as room temperature or 37°C, for up to 48 hours) to improve the nuclear localization and import efficiency of the vector.) The constructed vector enters the nucleus for expression and completes the insertion of the corresponding sequence into the corresponding site on the genome. By continuing genome insertion according to the above method based on the new site generated after insertion, sequence insertion can be performed continuously, and long fragment insertion into the genome can be achieved without significant length restrictions. II. DNA-mediated genomic sequence deletion technology, as shown in Figure 2 1. Deletion of any region in the genome: The sequence to be inserted in the vector designed using the above insertion method is changed to a specific sequence (within 2000 bp) upstream or downstream of the insertion point (target site) (within 100,000 bp). Using the DNA-, RNP-, or RNA-mediated insertion approach described in this invention, the sequence between two identical sequences can be removed with a certain degree of efficiency by homologous recombination after sequence insertion. (Selection: A sequence containing a recombination site (GCAGA[A / T]C, CCCA[C / G]GAC, or / and CCAGC) can be selected for insertion to improve the efficiency of subsequent homologous recombination.) 2. CNV End Deletion: CNV ends in cells or tissues are detected by sequencing and alignment (alignment of the junction between the gene sequence and the partial SINE sequence). The gene portion (within 2000 bp) of the CNV end to be processed and the 3' portion of the lasso that can form a portion of the downstream range (within 2000 bp) of this CNV end in the complete gene (possible downstream lasso sequences can be predicted or detected using the following method) are selected (or the 3' portion of the lasso can be replaced with a truncated sequence within 20,000 bp of the CNV end in the complete gene). Then, adjacent sequences (sequences) (immediately upstream sequences) upstream (within 100,000 bp) of the CNV end to be deleted are inserted. A complete SINE sequence, partial SINE sequence, or similar SINE sequence (using the different insertion techniques described above) is inserted (optionally, the ORF1p and ORF2p coding sequences can then be inserted). Next, the above sequences are synthesized to construct a vector. Then, using one of the gene insertion techniques described above, the upstream sequence immediately adjacent to the sequence to be deleted at the CNV end is inserted between the gene portion of the actual CNV end and the SINE sequence portion via the DNA, RNA, or RNP pathway (the SINE sequence used in the vector should be the same as or close to the SINE sequence surrounding the insertion site to increase efficiency). The sequence to be deleted is then deleted by homologous recombination between the same sequences (optionally, sequences containing recombination sites (GCAGA[A / T]C, CCCA[C / G]GAC, or / and CCAGC) can be selected for insertion to improve the efficiency of subsequent homologous recombination). III. DNA-mediated genome sequence replacement technology: The sequence inserted into a vector designed using insertion technology is modified to include the replacement sequence and the surrounding sequence of the sequence to be replaced in the genome (i.e., the sequence to be deleted by homologous recombination between the inserted replacement sequence and the corresponding sequence in the genome. When constructing the vector, the insertion site is located 3' or 5' of the replacement sequence, depending on whether it is upstream or downstream of the sequence to be replaced in the genome) (the replacement sequence must be homologous to the sequence to be replaced in the genome). The replacement sequence and the surrounding sequence of the sequence to be replaced in the genome are inserted upstream or downstream of the sequence to be replaced in the genome using gene editing insertion technology. When the inserted replacement sequence and the sequence to be replaced in the genome undergo homologous recombination, the sequence to be replaced in the genome is replaced by the homologously inserted replacement sequence. At the same time, the surrounding sequence of the deleted replacement sequence is reinserted into the genome by homologous recombination along with the sequence to be replaced during the sequence insertion. RNA-mediated genome sequence editing technology (because it does not require conversion of RNA to DNA and does not require the addition of additional TTAAAA or TTAAAA sites), as shown in Figure 3. 1. Ribonucleoprotein (RNP)-mediated pathway: The synthesized vector is amplified and transfected into an engineered cell line that highly expresses the protein encoded by the LINE (the efficiency can be increased by selecting a LINE that corresponds to the function of the SINE sequence contained in the synthetic vector; for example, if the synthetic vector contains an Alu sequence or a partial Alu sequence, it corresponds to LINE-1 and its encoded proteins ORF 1 p and ORF 2 p). (Engineered cells that permanently overexpress the relevant protein can be obtained by transfection with a vector expressing the relevant protein or post-transfection screening; if the ribonucleotides used in the subsequent step are incubated with the relevant protein, such as ORF 1 p and ORF 2 p, the engineered cells into which the transfection is performed do not need to express the relevant protein). After a period of time, the cell nuclei and cytoplasm are extracted, and the single-stranded plasmid product (single-stranded RNA) or a biologically active ribonucleoprotein (RNP) complex containing the single-stranded plasmid product (single-stranded RNA) is extracted using principles such as sequence specificity and corresponding conventional methods. (The resulting cytoplasm containing ORF 1 p and / or ORF 2 p or ORF 1 p is extracted.) Alternatively, the RNP complex can be incubated in physiological fluid containing ORF1p and / or ORF2p (at room temperature or 37°C for up to 48 hours) and then extracted and purified again. RNAse inhibitors must be added to the in vitro physiological fluid or cytoplasm, and if the transfected cells do not express the relevant proteins, incubation with ORF1p and / or ORF2p must be performed beforehand. The RNP complex can then be transferred into in vitro cultured cells or tissues using standard transfection methods such as liposomes or viral transfection (transfer to the cytoplasm eliminates the need for direct entry into the cell nucleus), or delivered to the living body via blood, lymph, cerebrospinal fluid, or local tissues to complete gene editing. If directional delivery is required, the outer wrapping of the vector can be modified. Note that the entire process avoids RNA degradation. Furthermore, when applying vectors synthesized using the LINE-mediated method in the RNP format, it is necessary to screen biologically active ribonucleoprotein (RNP) complexes containing extracted single-stranded plasmid products (single-stranded RNA) for products that do not contain the distal LINE-1 sequence or the ORF 1p and ORF 2p coding sequences (due to sequence specificity). (Additional in vitro nucleotidases can be added to promote shearing.) This is done to prevent the distal sequence from disrupting the gene editing target. 2. Simple RNA-mediated pathway: As described above, upstream and downstream sequences (each within 2000 bp) including the insertion point (i.e., the target site) and the sequence to be inserted (within 2000 bp) located in the middle corresponding to the insertion point are synthesized. Then, a SINE sequence, partial SINE sequence, or similar SINE sequence is connected, followed by the LINE sequence or the protein-coding sequence contained therein corresponding to the SINE function to be used (e.g., if a partial Alu sequence is used, it corresponds to the LINE-1 and ORF1 p and ORF2 p coding sequences therein; if a partial MIR sequence is used, it corresponds to the LINE-2 and the corresponding protein-coding sequence therein). This sequence is then constructed into a vector and driven by an RNA polymerase II / III promoter. (Alternatively, the carriers obtained by the various DNA-mediated methods described above can be directly transferred into engineered cells, and then the RNA product usable for gene editing is extracted by conventional means based on sequence specificity, etc.) After isolating and purifying the expressed mRNA, the RNA can be packaged using conventional transfection methods, such as liposomes or viruses, and then introduced into cells or tissues cultured in vitro (as long as it is delivered to the cytoplasm, not the nucleus) or into the body via blood, lymph, cerebrospinal fluid, or local tissue administration, achieving the goal of inserting the target sequence into the corresponding target site in the genome. By continuing genome insertion according to the above method based on the new site generated after insertion, sequence insertion can be performed continuously, allowing for the insertion of long fragments into the genome without significant length limitations (although it may be necessary to continuously transfect cells with RNA). It inhibits genomic changes caused by translocations and stabilizes the genome and its CNVs. (That is, this gene editing technology inserts a sequence that is non-cognate to the upstream and downstream sequences in the genome or the CNV terminal gene segment and the entire gene segment between the CNV terminal gene segment and the partial SINE sequence or in other regions, thereby inhibiting further CNV extension. The CNV terminal is defined as the region where the gene sequence (segment) is directly connected to the partial SINE sequence, where the gene can be extended. The specific sequences of each specific CNV terminal gene sequence (segment) and partial SINE sequence can be obtained by molecular biology methods such as gene sequencing or gene chips.) (The corresponding vector is packaged using conventional transfection methods such as lipid-soluble substances or substances with cell transfection capabilities, such as liposomes or viruses, and then transfects in vitro cultured cells or tissues, or is introduced into the body via blood, lymph, cerebrospinal fluid, or local tissue administration) (as shown in Figure 4). 1. Intervention in a specific CNV (a gene segment where the upstream sequence of the target site used to achieve insertion is the CNV terminus of a specific gene): Select the CNV to be manipulated, and use the boundary between the 3' end of the gene segment and the partial SINE sequence as the insertion point (target site). The 3' end (within 2000 bp) of the CNV terminus gene segment is used as the upstream sequence of the insertion point (target site) in the above insertion method. The downstream sequence of the target site is a partial SINE sequence (thus, SINE sequences, partial SINE sequences, or similar SINE sequences connected after the downstream sequence of the target site in the above method can be omitted). The inserted sequence is any sequence (within 2000 bp) non-homologous to the gene segment at the genome or CNV terminus or the upstream and downstream sequences of the gene segment in the complete gene. After vector construction is complete, it is transferred into the corresponding cell, tissue, or organism via the above-mentioned DNA, RNA, or RNP pathways, and the heterologous sequence is inserted into the corresponding CNV terminus. Because the non-homologous sequence does not exist downstream of the corresponding CNV terminal gene sequence (portion) in the complete gene, the CNV terminal cannot be further extended based on the complete gene sequence, thereby inhibiting further variation of the CNV terminal. 2. Intervention in widespread CNVs in the genome (the upstream sequence of the target site used to achieve insertional effect must include the gene portion of all CNV ends that may exist): (1) Genome disruption sequencing: Cells from the organism, tissue, or cell line requiring manipulation are cultured in vitro or directly extracted, and then sonicated and amplified using random primers and PCR. A short random sequence (20 bp or less) is designed and synthesized, and a partial SINE sequence is added downstream. The resulting genomic fragments and the synthesized short random sequence-connecting SINE sequence fragments are then connected and amplified by PCR to obtain the SINE sequence connecting the different genomic fragment sequences. The resulting fragments are then constructed into vectors and transferred into the corresponding cells, tissues, or organisms via the aforementioned DNA, RNA, or RNP pathways. All CNV ends in the genome are targeted via the genomic fragment sequences, and a non-homologous sequence (i.e., a short random sequence or a partial short random sequence that is not cognate to the gene fragment but is non-cognate to the local gene sequence of the corresponding gene fragment) is inserted between the gene segment at the CNV end and the partial SINE sequence. Because the non-homologous sequence is not present downstream of the corresponding CNV end gene segment in the complete gene, further alterations of the CNV end are prevented. (2) Random sequence method: A plasmid is constructed by adding a random sequence of an appropriate length (within 100 bp) (including all possible sequence combinations and excluding sequences similar to SINE sequences) to any non-cognate genomic sequence (within 2000 bp), followed by a partial SINE sequence. Alternatively, a plasmid is constructed by adding a random sequence (within 100 bp) to the SINE sequence, followed by a natural shearing site (for example, in the case of an Alu element transcription product, this site is located in the middle of the transcription product, allowing shearing to generate scAlu and partial Alu). A partial SINE sequence is then added to the random sequence transcribed by RNA polymerase II, followed by a vector (which can then be expressed as a lasso). This vector must contain a SINE sequence or be additionally transferred to another vector containing a SINE sequence. A LINE sequence or its protein-coding sequence corresponding to the SINE function can be added or additionally expressed downstream of the SINE sequence. It is then transferred into the corresponding cells, biological tissues or organisms through the above-mentioned DNA, RNP or RNA pathways, targets all CNV ends on the genome through random sequences, inserts non-homologous sequences into the corresponding CNV ends, and inhibits further changes in the CNV ends because the non-homologous sequences do not exist downstream of the corresponding CNV end gene sequence (part) in the complete gene. (3) By using the lariat end sequence method: All types of lariat (inserting a small random sequence (within 100 bp) that is not homologous to the genome into the SINE sequence, and then successfully excising the SINE sequence into a partial SINE sequence (i.e., the insertion position of this non-homologous sequence is downstream of the natural SINE shearing site, not located at the shearing site), and then constructing a plasmid that can express this modified SINE sequence, and transferring it from the corresponding target organism to amplified cells or a cell line of the corresponding species (taking the genome of the corresponding target species and dividing the whole genome into a relatively long (more than 200 bp) and overlapping sequence (10 Alternatively, the fragments can be cut into fragments (overlapping by 1 or more bp) and assembled into vectors. These fragments are then overexpressed by RNA polymerase II in in vitro cells of the corresponding species. After a certain period of time, the corresponding nucleic acids are extracted due to the sequence specificity of the non-homologous sequence inserted into the SINE sequence, and sequencing tests are performed to obtain sequence information for the various generated lassos connected to the partial SINE sequence containing the non-homologous sequence. Alternatively, or simultaneously, lasso sequences can be predicted based on the sequence rules for forming lassos in pre-mRNA (for example, they often end with AG), and all lasso sequences for that species or individual can be obtained. The 3' sequences of all lariat sequences (within 2000 bp) are taken and any sequence (within 2000 bp) non-homologous to the genome is connected to each of them, and then the SINE sequence (as described above, to increase efficiency, a "LINE sequence corresponding to the SINE function or its protein-coding sequence" can be connected to the end of it) is integrated into an expression vector (SINEs can also be expressed in a separate vector), expressed as lariat sequences, and connected to a partial SINE sequence generated by shearing of the SINE transcript in the cell; alternatively, the 3' sequences of all the obtained lariat sequences are each connected to any sequence (within 2000 bp) non-homologous to the genome, and then the connected partial SINE sequence (as described above, a LINE sequence corresponding to the SINE function or its protein-coding sequence can be connected to the end of it to increase efficiency) (it is preferable that the SINE sequence is the same as or similar to the SINE sequence of the gene to which the 3' sequence of the lariat connected to it is located), and a vector is constructed and expressed.It is transferred to the corresponding cell, tissue, or organism via the DNA, RNP, or RNA pathways described above, and edits the CNV ends throughout the entire genome. (4) SINE sequence modification: In other words, by administering the expression of a modified SINE sequence, a sequence non-homologous to the genome or gene portion in the CNV end or the upstream and downstream sequences of this portion in the complete gene is inserted into each CNV end to inhibit CNV end extension. A short sequence (a short sequence (within 100 bp) that does not match the 3' sequence of the socket normally generated and spans the SINE natural shearing site) is added before the SINE natural shearing site, and a complete SINE sequence is constructed to enable natural shearing of the SINE transcript in this new region (efficiency can be increased by adding a LINE sequence or its protein-coding sequence corresponding to the corresponding type of SINE function after it), or a complete SINE sequence is constructed in which any genomic non-homologous sequence (within 200 bp) is added after the natural shearing site of the SINE transcript (efficiency can be increased by adding a LINE sequence or its protein-coding sequence corresponding to the corresponding type of SINE function after it), and then the corresponding cells, tissues, or organisms are administered. The SINE sequences used should cover as many SINE sequences (which can be obtained by methods such as sequencing or array chips) of the species or individual as possible in order to accurately correct all CNV ends across the entire genome. The entire genome can also be cut into long fragments that overlap each other to a certain extent (with an overlapping length equal to or greater than the length of a single lariat structure), which can then be incorporated into a vector and overexpressed in an in vitro cell line of the corresponding species to produce lariat structures. The vector containing the modified SINE sequence (which can then be mediated through the RNA pathway after adding a LINE sequence or its protein-coding sequence corresponding to the corresponding type of SINE function downstream) is then transferred into the vector, and the single-stranded RNA ribonucleoprotein (RNP) complex or RNA with the biological activity of the partial SINE (originating from the modified SINE sequence) connected to the lariat (derived from the modified SINE sequence) is isolated and purified using sequence specificity and other properties and conventional means, and then functions through the corresponding RNA or RNP pathway. 3. Modifying SINEs and LINEs in the genome: Using this gene editing technology, any sequence (up to 500 bp) can be inserted into the promoter, natural shearing site of the transcript, or other sequence of a SINE in the genome, or / and the promoter, protein-coding sequence, or other sequence of a LINE, so that the SINE sequence in the genome cannot be transcribed or sheared after transcription, and / or the LINE sequence in the genome cannot be transcribed or cannot produce a functional protein. First, the SINE or LINE sequence in the entire genome of the individual to be manipulated is sequenced, and the promoter, natural shearing site of the transcript, protein-coding sequence, or other sequence therein is selected as the insertion point (target site). In this gene editing technology, the upstream and downstream sequences of the insertion site (target site) on the SINE or LINE sequence are those upstream and downstream of the insertion site (target site) on the SINE or LINE sequence, and the inserted sequence can be any sequence. Using the above insertion method, any sequence can be inserted into the corresponding site on the SINE or LINE in the genome. Alternatively, the SINE or LINE sequence in the genome can be inactivated by replacing or deleting it using the above gene editing method. 4. Fixing the CNV end while deleting it: Select the CNV end that requires manipulation, and use the boundary between the 3' end of the gene portion and the partial SINE sequence as the insertion site. In the insertion method, the upstream sequence of the target site is the 3' end sequence of the gene portion of the CNV end (within 2000 bp), and the downstream sequence is the partial SINE sequence (therefore, the partial SINE sequence connected after the downstream sequence of the target site in the above gene editing method can be omitted). The sequence to be inserted is the sequence (within 2000 bp) adjacent upstream of the sequence to be deleted on the genome (within 100,000 bp), followed by any sequence (within 2000 bp) that is non-homologous to the genomic sequence. After the vector construction is complete, it is transferred into the corresponding cells, tissues, or organisms via the above-mentioned DNA, RNA, or RNP pathways, and the sequence adjacent to the sequence to be deleted on the genome and the subsequent non-homologous sequence are inserted into the corresponding CNV end. After homologous recombination occurs between the two identical (homologous) sequences to delete the intermediate sequence, the non-homologous sequence simultaneously inhibits further extension of the CNV. 5. Intrinsic mechanism suppression method: The CNV elongation mechanism inherent to cells or organisms can also be directly suppressed. For example, RNA interference can be used to suppress the transcription of SINEs and LINEs, as well as the production of their RNA and encoded proteins, such as ORF 1p and ORF 2p. Specific proteins can then be used to bind to the functional structures of proteins associated with this CNV elongation mechanism, such as ORF 1p, ORF 2p, or shear bodies or complexes, thereby inhibiting their function. The above-mentioned gene editing techniques can be used to modify and inactivate or reduce the activity of SINEs, such as Alu and various MIRs, various LINEs, and their corresponding protein-coding sequences in the genome, thereby suppressing the function of proteins involved in homologous recombination or DNA mismatch repair, or to administer modified nucleoside compounds to inhibit reverse transcription. This suppression of the endogenous CNV elongation mechanism can thereby inhibit genomic changes and stabilize CNVs. Because SINEs, LINEs, and the proteins they express are widely distributed in eukaryotes, this technology can be used to perform gene editing on a wide range of eukaryotes, and can also be applied to the treatment of diseases caused by genetic modification and the modification or stabilization of cellular or biological states related to genetic modification. In this technique, a determined sequence (such as a sequence to be inserted) is defined as being in the 5'→3' direction, with upstream being before the 5' end of the determined sequence and downstream being after the 3' end of the determined sequence, with the upstream sequence being the sequence before the 5' end of the determined sequence and the downstream sequence being the sequence after the 3' end of the determined sequence. As shown in Figure 5, the gene transcription framework provided by this technology comprises a target site upstream sequence, an insert sequence (sequence to be inserted), and a target site downstream sequence in the 5'→3' direction. To better understand the positional relationship between the gene transcription framework, short interspersed elements, long interspersed elements, and promoter, several different connection configurations are presented. Figure 6 is a schematic diagram of a structure in which a promoter is linked to the front of the gene transcription framework. The promoter may be an RNA polymerase I promoter, an RNA polymerase II promoter, or an RNA polymerase III promoter. The promoter can be located on a vector, and the gene transcription framework, short interspersed elements, long interspersed elements, etc. are inserted downstream of the promoter through a vector enzyme cleavage site, followed by transfection into cells for subsequent expression. A direct synthesis method can also be used to directly synthesize the gene transcription framework, short interspersed elements, long interspersed elements, etc., and insert them into a vector. Figure 7 shows a structure in which a promoter is connected upstream of the gene transcription framework and short interspersed elements, partial short interspersed elements, or similar short interspersed elements are connected downstream. Figure 8 is a structural diagram of a gene transcription framework in which a promoter is connected upstream and a long interspersed nuclear element or ORF1p coding sequence or ORF2p coding sequence is connected downstream. Figure 9 is a structural diagram of a gene transcription framework in which a long interspersed nuclear element or ORF1p coding sequence or ORF2p coding sequence is connected upstream and a promoter is connected upstream of the long interspersed nuclear element or ORF1p coding sequence or ORF2p coding sequence. Figure 10 is a structural diagram of a gene transcription framework in which a promoter is connected upstream and a short interspersed nuclear element, partial short interspersed nuclear element, or similar short interspersed element is connected downstream, followed by a long interspersed nuclear element or ORF1p coding sequence or ORF2p coding sequence.Figure 11 is a schematic diagram of a structure in which a short interspersed nuclear element, a partial short interspersed nuclear element, or a similar short interspersed nuclear element is connected downstream of a gene transcription framework, a long interspersed nuclear element or ORF1p coding sequence or ORF2p coding sequence is connected upstream of the gene transcription framework, and a promoter is connected upstream of the long interspersed nuclear element, ORF1p coding sequence, or ORF2p coding sequence. Figure 12 is a schematic diagram of a structure in which the gene transcription framework and the short interspersed nuclear element, a partial short interspersed nuclear element, or a similar short interspersed nuclear element do not share a single promoter. Figure 13 is a schematic diagram of a structure in which the gene transcription framework and the long interspersed nuclear element, ORF1p coding sequence, or ORF2p coding sequence do not share a single promoter. Figure 14 shows a gene transcription framework and a short interspersed nuclear element, a partial short interspersed nuclear element, or a similar short interspersed element, and a long interspersed nuclear element, or an ORF1p coding sequence or ORF2p coding sequence, but they do not share a single promoter structure; the short interspersed nuclear element, a partial short interspersed nuclear element, or a similar short interspersed element is located downstream of the long interspersed nuclear element, or an ORF1p coding sequence or ORF2p coding sequence, and they share a single promoter. Figure 15 shows a gene transcription framework and a short interspersed nuclear element, a partial short interspersed nuclear element, or a similar short interspersed element, and a long interspersed nuclear element, or an ORF1p coding sequence or ORF2p coding sequence, but they do not share a single promoter; the short interspersed nuclear element, a partial short interspersed nuclear element, or a similar short interspersed element is located upstream of the long interspersed nuclear element, or an ORF1p coding sequence or ORF2p coding sequence, and they share a single promoter. In all of these cases, the gene transcription framework and short interspersed nuclear elements, partial short interspersed nuclear elements, or similar short and long interspersed nuclear elements, or the ORF1p coding sequence or ORF2p coding sequence, are on the same vector, but do not have to be on the same vector; they may be co-transfected into cells and expressed by different promoters. In the following examples, the materials used are human-derived cells, and therefore the SINEs used are primate-specific short interspersed nuclear elements (Alu elements). The complete sequence of the Alu element is shown in Seq ID No. 1, and a partial Alu sequence is shown in Seq ID No. 2. If the materials used are from other species, the short interspersed nuclear elements can be exchanged for the corresponding species' short interspersed nuclear elements to facilitate expression. material 1. The pSIL-eGFP plasmid vector was Plasmid 52675 purchased from Addgene, and the pBS-L1PA1-CH-mneo plasmid vector was Plasmid 51288 purchased from Addgene. 2. 10x enzyme cleavage buffer (required for NheI enzyme cleavage): 330 mM Tris-acetate, 100 mM magnesium acetate, 660 mM potassium acetate, 1 mg / mL BSA; 10x enzyme cleavage buffer (required for Sali enzyme cleavage): 500 mM Tris-HCl, 100 mM MgCl2, 1000 mM NaCl, 1 mg / mL BSA. 3. Restriction enzymes NheI and Sali were purchased from ThermoFisher. 4. T4 DNA ligase and the 10x ligation buffer required for its application were purchased from Promega. 5. Entranster-H 4000 transfection reagent was purchased from Beijing Yinggeon Biotechnology Co., Ltd. 6. Blood / cell / tissue genomic DNA extraction kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., product catalog number: DP 304. 7. SuperReal PreMix Plus (SYBR Green) was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., product catalog number: FP205. 8. Magnetic bead method tissue / cell / blood total RNA extraction kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., product catalog number: DP 761. 9. FastKing cDNA First Strand Synthesis Kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., product catalog number: KR 116. 10. TIANSeq mRNA capture kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., product catalog number: NR 105. 11. Lipofectamine TM MessengerMAX TM mRNA transfection reagent was purchased from ThermoFisher. 12. Trypsin was purchased from Sigma-Aldrich, product catalog number: T 1426. 13. Dnase I was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., product catalog number: RT411. 14. The chemical synthesis of primers and sequences was completed by Poshang Biotechnology (Shanghai) Co., Ltd. Example 1 DNA-mediated insertion of exogenous sequences into specific genomic sites Vascular endothelial growth factor (VEGFA) is a member of the PDGF / VEGF growth factor family. It encodes a heparin-binding protein that exists as a disulfide-linked homodimer. This growth factor plays essential roles in both physiological and pathological angiogenesis, including angiogenesis, endothelial cell proliferation, induction of endothelial cell proliferation, promotion of cell migration, inhibition of apoptosis, and induction of vascular permeability. In this example, an exogenous sequence was inserted into the VEGFA gene to verify the DNA-mediated genomic sequence insertion technique of the present invention. A 459-bp sequence of the gene VEGFA in the human genome was selected and is shown in SEQ ID NO:3: ATTATGCGGATCAAACCTCACCAAGGCCAGCACATAGGAGAGATGAGCTTCCTACAGCACAACAAATGTGAATGCAGGTGAGGATGTAGTCACGGATTCATTATCAGCAAGTGGCTGCAGGGTGCCTGATCTGTGCCAGGGTTAAGCATGCTGTACTTTTTGGCCCCCGTCCAGCTTCCCGCTATGTGACCTTTGGCATTTTACTTCAATGTGCCTCAGTTTCTACATCTGTAAAATGGGCA C*AATAGTAGTATACTTCATAGCATTGTTATAATGATTAAACAAGTTATATGAAAAGATTAAAACAGTGTTGCTCCATAATAAATGCTGTTTTTACTGTGATTATTATTGTTGTTATCCCTATCATTATCATCACCATCTTAACCCTTCCCTGTTTTGCTCTTTTCTCTCTCCCTACCCATTGCAGACCAAAGAAAGATAGAGCAAGACAAGAAAA, where * was the selected insertion site (target site). The upstream sequence of the insertion site in the VEGFA gene (target site upstream sequence) preceded the insertion site, and the downstream sequence of the insertion site in the VEGFA gene (target site downstream sequence) followed the insertion site. A randomly designed heterologous sequence was added to the insertion site to form the gene transcription framework, and restriction endonuclease NheI enzyme cleavage sites and protective bases (overhangs) were added to both ends of the gene transcription framework to enable insertion of the gene transcription framework into an expression vector. The complete sequence is shown in SEQ ID NO:4: CTAGCTAGCTAGATTATGCGGATCAAACCTCACCAAGGCCAGCACATAGGAGAGATGAGCTTCCTACAGCACACAACAAATGTGAATGCAGGTGAGGATGTAGTCACGGATTCATTATCAGCAAGTGGC TGCAGGGTGCCTGATCTGTGCCAGGGTTAAGCATGCTGTACTTTTTGGCCCCCGTCCAGCTTCCCGCTATGTGACCTTTGGCATTTTACTTCAATGTGCCTCAGTTTCTACATCTGTAAAATGGGCAC GCTACGGAATAAGAGGAGGCCACAACAGTCGTGGGTCGAATAGTAGTATACTTCATAGCATTGTTATAATGATTAAACAAGTTATATATGAAAAGATTAAAACAGTGTTGCTCCATAATAAATGCTGTTTTTACTGTGATTATTATTGTTGTTATCCCTATCATTATCATCACCATCTTAACCCTTCCCTGTTTTGCTCTTTTCTCTCTCCCTACCCATTGCAGACCAAAGAAAGATAGAGCAAGACAAGAAAACTAGCTAGCTAG, where the underlined portion is the randomly designed exogenous non-homologous sequence (i.e., the sequence to be inserted). It is 38 bp long, and both ends of the sequence are NheI enzyme cleavage sites and protecting bases (bold italics). The sequence was obtained by chemical synthesis and named VEGFA1. The exogenous sequence to be inserted was designed as a non-homologous sequence to that of the VEGFA gene, so that insertion into the target site on the genome could be specifically detected in subsequent experiments. At the same time, short sequences upstream and downstream of the VEGFA gene insertion site were added before and after the insertion site on the vector to verify the effect of different lengths of upstream and downstream sequences of the target site on the insertion effect. The insertion site of the VEGFA sequence was selected 10 bp before and 10 bp after the insertion site, and a sequence having a non-homologous sequence, an NheI enzyme cleavage site and a protective base was designed into a short sequence as shown in SEQ ID NO: 5: CTAGCTAGCTAGAAATGGGCAC GCTACGGAATAAGAGGAGGCCACAACAGTCGTGGGTCG AATAGTAGTACTAGCTAGCTAG, where the underlined part represents a randomly designed 38-bp exogenous non-homologous sequence (the sequence to be inserted), with NheI enzyme cleavage sites and protective bases (bold italics) at both ends of the sequence. The sequence obtained by chemical synthesis was named VEGFA2. The choice of restriction enzyme cleavage site was solely for convenience of plasmid construction and could be varied for different vectors. The specific process of inserting VEGFA1 and VEGFA2 into pSIL-eGFP to construct the plasmids pSIL-eGFP-VEGFA1 and pSIL-eGFP-VEGFA2 is as follows: VEGFA1, VEGFA2, and pSIL-eGFP were digested separately, and the reaction system is shown in Table 1: Table 1. Digestion reaction system The reaction conditions were incubation at 37°C for 1 hour, followed by incubation at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested VEGFA1 and VEGFA2 were ligated with the plasmid vector pSIL-eGFP, and the reaction system is shown in Table 2: Table 2. Linkage reaction system The reaction was incubated at 16°C for 16 hours, followed by 10 minutes at 70°C to inactivate the ligase. The plasmids pSIL-eGFP-VEGFA1 and pSIL-eGFP-VEGFA2 were purified by electrophoresis and sequenced to confirm the integrity of the plasmids. The pSIL-eGFP plasmid has its own CMV promoter, so transcription can be initiated by RNA polymerase II as long as the expression framework is inserted into the CMV promoter. An Alu expression sequence is designed, and the Alu sequence, the heterologous sequence (18 bp), TTTTT, and TTTTAA*n (where n is 6) are ligated, and SalI enzyme cleavage sites and corresponding protecting bases are added to both ends of the sequence to obtain the sequence shown in SEQ ID NO:6: ACGCGTCGACGTCGGCCATAGCGGCCGCGGAAGGGCCGGGCGCGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGGCGGATCACGAGGTCAGGAGATCGAGACCATCCCGGCTAAAACGGTGAAACCCCGTCTCTACTAAAAATACAAAAAATTAGCCGGGCGTGGTGG CGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCCCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATC CTATCAGGCGAGCCCGTATTTTTTTTTAATTTTAATTTTAATTTTAATTTTAATTTTAA The sequence is ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA, where the italics at both ends represent the bolded SalI digestion site and corresponding protected bases. The upstream SalI digestion site and corresponding protected base are followed by the Alu sequence. The underline represents the non-homologous sequence (18 bp) added to Alu. This is used to label Alu and facilitate detection of its transcript binding with the lasso generated by transcription of the gene transcription framework (including the inserted sequence) in later experiments to verify the mechanism of action of the experiment. The wavy line represents the transcription terminator, and the double underline represents a 6TTTTAA repeat used to convert RNA to DNA, a common repeating sequence in genomes. You can choose to add or not add multiple sequences; in this embodiment, we chose to add 6. The sequence was obtained by chemical synthesis and designated Alu1. Alu1, plasmid pSIL-eGFP-VEGFA1, and plasmid pSIL-eGFP-VEGFA2 were each digested with SalI. The digestion reaction system is shown in Table 3: Table 3. Digestion reaction system The reaction conditions were incubation at 37°C for 3 hours, followed by incubation at 80°C for 10 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested Alu1 was ligated with the plasmid vectors pSIL-eGFP-VEGFA1 and pSIL-eGFP-VEGFA2, and the reaction system is shown in Table 4: Table 4. Linkage reaction system The reaction conditions were incubation at 16°C for 16 hours, followed by incubation at 70°C for 10 minutes to inactivate the ligase. The resulting products were electrophoresed and purified to obtain the plasmids pSIL-eGFP-VEGFA1-Alu1 (shown in Figure 16) and pSIL-eGFP-VEGFA2-Alu1. The plasmids were sequenced to confirm their authenticity. The pSIL-eGFP plasmid itself contains the U6 promoter, an RNA polymerase III-dependent promoter. Therefore, if an Alu sequence is inserted after the U6 promoter, transcription of the Alu sequence (Alu element) may be initiated by RNA polymerase III. pSIL-eGFP-VEGFA1-Alu1 or pSIL-eGFP-VEGFA2-Alu1 was transfected into HeLa cells to test the insertion efficiency of the randomly designed exogenous sequences. To improve the insertion efficiency, pBS-L1PA1-CH-mneo, a plasmid expressing ORF1p and ORF2p (LINE), was co-transfected into HeLa cells, and a corresponding control group was designed. The co-transfected plasmids for the experimental and control groups are listed in Table 5. Table 5 Grouping As shown in the groupings, control group 1 consisted of co-transfected pSIL-eGFP and pBS-L1PA1-CH-mneo, which lacked the gene transcription framework sequence and Alu1 sequence. Experimental group 1 consisted of co-transfected pSIL-eGFP-VEGFA1-Alu1 and pBS-L1PA1-CH-mneo, which contained a gene transcription framework with long sequences upstream and downstream of the target site in the VEGFA gene, the Alu1 sequence, and pBS-L1PA1-CH-mneo. Experimental group 2 consisted of co-transfected pSIL-eGFP-VEGFA1 and pBS-L1PA1-CH-mneo, which contained a gene transcription framework with long sequences upstream and downstream of the target site in the VEGFA gene, but lacked the Alu1 sequence. In experimental group 3, pSIL-eGFP-VEGFA2-Alu1 and pBS-L1PA1-CH-mneo were co-transfected, containing a gene transcription framework with short sequences upstream and downstream of the target site in the VEGFA gene, an Alu1 sequence, and pBS-L1PA1-CH-mneo. In experimental group 4, pSIL-eGFP-VEGFA1-Alu1 was transfected, but pBS-L1PA1-CH-mneo was not. This vector contained a gene transcription framework with long sequences upstream and downstream of the target site in the VEGFA gene, and an Alu1 sequence, but pBS-L1PA1-CH-mneo did not. Each group contained three parallel 6-well plates cultured with HeLa cells. For the transfection procedure, HeLa cells were passaged and spread into 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each plate of cells, 48 ​​μg or 96 μg of the constructed plasmid (48 μg if only one plasmid was transfected, according to the experimental group; 48 μg of each plasmid was used if two plasmids were co-transfected, for a total of 96 μg of the two plasmids) was diluted with 300 μL of serum-free DMEM and mixed thoroughly. At the same time, 120 μL of Entranster-H4000 reagent was diluted with 300 μL of serum-free DMEM, mixed thoroughly, and allowed to stand at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and allowed to stand at room temperature for 15 minutes to create the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM culture medium containing 10% fetal bovine serum per well for transfection. When the cells are about 90% confluent, the above procedure is repeated after passage until the cells are about 90% confluent and then removed for the subsequent procedure. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for detecting the GAPDH gene is shown in SEQ ID NO: 7 as follows: 5'-CACTGCCACCCAGAAGACTG-3'. The downstream primer sequence is shown in SEQ ID NO: 8 as follows: 5'-CCTGCTTCACCACCTTCTTG-3'. Primer pair 1 and primer pair 2 were designed. The upstream primer sequence of primer pair 1 is shown as SEQ ID NO: 9: 5'-CCCAGGGTTGTCCCATCT-3'; the downstream primer sequence is shown as SEQ ID NO: 10: 5'-CCTCCTCTTATTCCGTAGC-3'. The upstream primer sequence of primer pair 1 is located in the complete VEGFA gene, and the upstream sequence of the insertion site (target site) used on the plasmid is located only in the genome, not within the plasmid. The downstream primer sequence of primer pair 1 is located in the 19-bp sequence at the 5' end of the randomly designed non-homologous sequence to be inserted (the inserted sequence). The upstream sequence of primer pair 2 is shown as SEQ ID NO: 11: 5'-CACAACAGTCGTGGGTCG-3'; the downstream primer sequence is shown as SEQ ID NO: 12: 5'-GAGGGAGAAGTGCTAAAGTCAG-3'. The upstream primer sequence of primer pair 2 is located in the 18-bp sequence at the 3' end of the randomly designed non-homologous sequence (the sequence to be inserted), and the downstream primer sequence is located in the entire VEGFA gene. The downstream sequence of the insertion site (target site) used on the plasmid is located only in the genome, not in the plasmid. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 6. Table 6 qPCR reaction system JPEG2024504592000007.jpg80107 Cell DNA templates were extracted from the control group 1 and experimental groups 1-4 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 1: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 50°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. Primer pair 2: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 54°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. The exponential growth phase was observed in the amplification curves for the detection of insertion of GAPDH and the inserted sequence, and it was confirmed that they were nearly parallel. -ΔΔCt The results are shown in Tables 7 and 8. Table 7 shows the results for primer pair 1, and Table 8 shows the results for primer pair 2. The PCR products were confirmed to be correct by sequencing. Table 7 Results of primer pair 1 (n = 3, x ± s) Compared with other groups (N / A calculated according to 40.00), the relative copy number in experimental group 1 was significantly higher than the other groups, with statistical significance (P < 0.05). This suggests that gene editing is most efficient due to longer upstream or downstream sequences of the insertion site (target site) and appropriate expression of Alu elements (SINEs) and / or ORF1p and / or ORF2p (LINEs) in the gene transcription framework. The relative copy number amounts in experimental groups 2, 3, and 4 were higher than in control group 1 (N / A calculated according to 40.00). All results were statistically significant (P < 0.05), suggesting that gene editing is still effective but less efficient. This is possible under the following conditions: the upstream or downstream sequence of the insertion site (target site) in the gene transcription framework is short, the cell itself has low or no expression of Alu elements (SINEs), and ORF1p and ORF2p (LINEs). Sufficient sequences on either side of the insertion site (target site) in the gene transcription framework are required to facilitate lariat formation to ensure efficient insertion. Editing efficiency can be improved by lengthening the upstream and / or downstream sequence of the target site or by additionally expressing ORF1p, ORF2p (LINEs), and / or Alu elements (SINEs). Table 8 Results of primer pair 2 (n=3, x±s) Compared with other groups (N / A calculated according to 40.00), the relative copy number in experimental group 1 was significantly higher than the other groups, with statistical significance (P < 0.05). This suggests that gene editing is most efficient due to longer upstream or downstream sequences of the insertion site (target site) and appropriate expression of Alu elements (SINEs) and / or ORF1p and / or ORF2p (LINEs) in the gene transcription framework. The relative copy number amounts in experimental groups 2, 3, and 4 were higher than control group 1 (N / A calculated according to 40.00). All results were statistically significant (P < 0.05), suggesting that gene editing is still effective but less efficient. This is possible under the following conditions: the upstream or downstream sequence of the insertion site (target site) in the gene transcription framework is short, the cell itself has low or no expression of Alu elements (SINEs), and ORF1p and ORF2p (LINEs). Sufficient sequences on either side of the insertion site (target site) in the gene transcription framework are required to facilitate lariat formation to ensure efficient insertion. Editing efficiency can be improved by lengthening the upstream and / or downstream sequence of the target site or by additionally expressing ORF1p, ORF2p (LINEs), and / or Alu elements (SINEs). The results in Tables 7 and 8 show that both ends of the inserted sequence were inserted into the target site on the genome, meaning that the inserted sequence was fully inserted into the target site. Experimental Groups 1, 2, 3, and 4 were able to fully insert the non-homologous sequence into the genome, with Experimental Group 1 being the most efficient. Example 2: Detecting connections between a lariat structure formed by a gene transcription framework (containing an exogenous sequence to be inserted) and an RNA fragment containing a SINE subsequence (e.g., an Alu element) (transcription of an Alu element cleaved at a natural shearing site) Extraction of total RNA from transfected cells in Experimental Group 1 and Control Group 1 in Example 1: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Transfer the cell-containing solution to an RNase-free centrifuge tube and centrifuge at 300 g for 5 minutes. Collect the pellet and aspirate all of the supernatant. Perform total RNA extraction according to the instructions of the magnetic bead method tissue / cell / blood total RNA extraction kit. Reverse transcription synthesis of cDNA template: The cDNA is synthesized according to the instructions of the FastKing cDNA First Strand Synthesis Kit, and the concentration of the synthesized cDNA is determined by UV spectrophotometer for subsequent testing. qPCR testing: The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 3 was designed to detect the junction between the transcriptional lariat structure containing the exogenous sequence to be inserted and the RNA fragment containing the partial Alu sequence produced by transcription of the Alu element. The upstream primer sequence is shown in SEQ ID NO:11: 5'-CACAACAGTCGTGGGTCG-3'. The upstream primer is located on the exogenous sequence to be inserted, and the downstream primer sequence is shown in SEQ ID NO:13: 5'-TACGGGCTCGCCTGATAG-3'. The downstream primer is located in the non-homologous sequence (18 bp) following the Alu sequence constructed in the plasmid. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 9. Table 9. qPCR reaction system JPEG2024504592000012.jpg80107 Reactions were prepared on ice. After preparation, the reaction tubes were capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 54°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. The exponential growth cycles in the amplification curves of GAPDH and the binding between the lariat structure containing the inserted sequence and the RNA fragment containing the partial Alu were observed and confirmed to be nearly parallel. -ΔΔCt The results are shown in Table 10. The PCR products were confirmed to be correct by sequencing. Table 10 Results of primer pair 3 (n=3, x±s) Compared with control group 1 (N / A calculated according to 40.00), the relative copy number in experimental group 1 was significantly higher with statistical significance (P < 0.05), indicating that the lariat construct containing the inserted exogenous sequence was indeed linked to the Alu sequence transcript (i.e., the RNA fragment containing the partial Alu). Table 10 shows that the transcripts of the Alu sequences (i.e., RNA fragments containing partial Alu) were indeed linked to the transcripts containing the inserted sequences. Example 3. Insertion of exogenous sequences into specific genomic sites via DNA The MMP2 gene, a member of the matrix metalloproteinase (MMP) gene family, is a zinc-dependent enzyme capable of cleaving extracellular matrix components and molecules involved in signal transduction. The protein encoded by this gene, collagenase A, type IV collagenase, contains three fibronectin type II repeats in the catalytic site, allowing it to bind to denatured types IV and V collagen and elastin. Unlike most MMP family members, activation of this protein can occur on the cell membrane. This enzyme can be activated extracellularly by proteases or intracellularly via S-glutathione without requiring proteolysis to remove the protodomain. This protein is thought to be involved in various pathways, including the nervous system, endometrial rupture, angiogenesis regulation, and metastasis. Mutations in this gene are associated with Winchester syndrome and tuberous arthropathy osteolysis (NAO) syndrome. Alternative splicing results in the encoding of multiple transcript variants with distinct subtypes. In this example, an exogenous sequence was inserted into the MMP2 gene to verify the DNA-mediated genomic sequence insertion technique of the present invention. A 479-bp sequence of the gene MMP2 in the human genome was selected and the sequence is shown in SEQ ID NO: 14: AGCATGGCGATGGATACCCCTTTGACGGTAAGGACGGACTCCTGGCTCATGCCTTCGCCCCAGGCACTGGTGTTGGGGGAGACTCCCATTTTGATGACGATGAGCTATGGACCTTGGGAGAAGGCCAAGGTGAGAAAGGGGCCCTCTGCATGCCCCAGACCTTCTCTCCTGTCCTCTCTCCACTCCATTTGCTTGGACCAGAGA G*GTGGGAGGGGAGGAAAGTCACACATCTGGGTGAGTCAGAATCTTGGTCTCCAAAGAAGGCCTGGAGAAGTCCAACCTCCCCCTTCCATGTCACTCTTTAGTGGTCCGTGTGAAGTATGGGAACGCCGATGGGGAGTACTGCAAGTTCCCCTTCTTGTTCAATGGCAAGGAGTACAACAGCTGCACTGATACCGGCCGCAGCGATGGCTTCCTCTGGTGCTCCACCACCTACAACTTTGAGAAGGATGGCAAGTACGGCTTCTGTCCCCATGAAG, where * was the selected insertion site (target site). The upstream sequence of the insertion site in the MMP2 gene (target site upstream sequence) was before the insertion site, and the downstream sequence of the insertion site in the MMP2 gene (target site downstream sequence) was after the insertion site. The sequence inserted to form the gene transcription framework was a randomly designed heterologous sequence added to the insertion site, and restriction endonuclease NheI enzyme cleavage sites and protective bases (overhangs) were added to both ends of the gene transcription framework to enable the gene transcription framework to be inserted into an expression vector. The complete sequence is shown in SEQ ID NO: 15: CTAGCTAGCTAGAGCATGGCGATGGATACCCCTTTGACGGTAAGGACGGACTCCTGGCTCATGCCTTCGCCCCAGGCACTGGTGTTGGGGGAGACTCCCATTTTGATGACGATGAGCTATGGACCTTGGGAGAAGGCCAAGGTGAGAAAGGGGCCCTCTGCATGCCCCAGACCTTCTCTCCTGTCCTCTCTCCACTCCATTTGCTTGGACCAGAGAG CCTGTGGGCCTTGCTCAGAGCGGAGAAAGCATGGCATAATGATGTGGCTGTTTTGTTTGTACAAGATCCGCAGACGTGTAAATGTTCCTGCAAAAACACAGACThe sequence is: GTGGGAGGGGAGGAAAGTCACACATCTGGGTGAGTCAGAATCTTGGTCTCCAAAGAAGGCCTGGAGAAGTCCAACCTCCCCCTTCCATGTCACTCTTTAGTGGTCCGTGTGAAGTATGGGAACGCCGATGGGGAGTACTGCAAGTTCCCCTTCTTGTTCAATGGCAAGGAGTACAACAGCTGCACTGATACCGGCCGCAGCGATGGCTTCCTCTGGTGCTCCACCACCTACAACTTTGAGAAGGATGGCAAGTACGGCTTCTGTCCCCATGAAGCTAGCTAGCTAG, where the underlined portion is the randomly designed exogenous non-homologous sequence (i.e., the sequence to be inserted), is 103 bp in length, and both ends of the sequence are NheI enzyme cleavage sites and protecting bases (bold italics). The sequence was obtained by chemical synthesis and named MMP2-1. The exogenous sequence to be inserted is designed to be non-homologous to that sequence of the MMP2 gene, so that insertion into the target site on the genome can be specifically detected in subsequent experiments. At the same time, short sequences upstream and downstream of the MMP2 gene insertion site were added before and after the insertion site on the vector to verify the effect of different lengths of upstream and downstream sequences of the target site on the insertion effect. 10 bp before and after the insertion site of the MMP2 sequence were selected, and a sequence having a non-homologous sequence, an NheI enzyme cleavage site and a protective base was designed into a short sequence as shown in SEQ ID NO: 16: CTAGCTAGCTAGGACCAGAGAG CCTGTGGGCCTTGCTCAGAGCGGAGAAAGCATGGCATAATGATGTGGCTGTTTTGTTTGTACAAGATCCGCAGACGTGTAAATGTTCCTGCAAAAACACAGAC GTGGGAGGGGCTAGCTAGCTAG, where the underlined part represents a randomly designed 103-bp exogenous non-homologous sequence (the sequence to be inserted), with NheI enzyme cleavage sites and protective bases (bold italics) at both ends of the sequence. The sequence obtained by chemical synthesis was named MMP2-2. The choice of restriction enzyme cleavage sites is for convenience of plasmid construction only and may vary depending on different vectors. After inserting MMP2-1 and MMP2-2 into pSIL-eGFP to construct the plasmids pSIL-eGFP-MMP2-1 and pSIL-eGFP-MMP2-2, the specific process is as follows: pSIL-eGFP, MMP2-1 and MMP2-2 were digested separately, and the reaction system is shown in Table 11: Table 11 Digestion reaction system The reaction was incubated at 37°C for 1 hour, then at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested MMP2-1 and MMP2-2 were ligated with the plasmid vector pSIL-eGFP, and the reaction system is shown in Table 12: Table 12. Linkage reaction systems The reaction was incubated at 16°C for 16 hours, followed by 10 minutes at 70°C to inactivate the ligase. The plasmids pSIL-eGFP-MMP2-1 and pSIL-eGFP-MMP2-2 were purified by electrophoresis and sequenced to confirm the integrity of the plasmids. The pSIL-eGFP plasmid has its own CMV promoter, so transcription can be initiated by RNA polymerase II as long as the expression framework is inserted into the CMV promoter. The Alu expression sequence was designed and consisted of the Alu sequence, the non-homologous sequence (18 bp), TTTTT, and TTTTAA*n (where n is ACGCGTCGACGTCGGCCATAGCGGCCGCGGAAGGGCCGGGCGCGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGGCGGATCACGAGGTCAGGAGATCGAGACCATCCCGGCTAAAACGGTGAAACCCCGTCTCTACTAAAAATACAAAAAATTAGCCGGGCGTGGTGG CGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCCCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATC CAAGAGCGAGCCAACAGATTTTTTTTTAATTTTAATTTTAATTTTAATTTTAATTTTAA The sequence is ACGCGTCGACGTCGGCCATAGCGGCCGCGGAA, where the italics at both ends represent the bolded SalI digestion site and corresponding protected bases. The upstream SalI digestion site and corresponding protected base are followed by the Alu sequence. The underline represents the non-homologous sequence (18 bp) added to Alu. This is used to label Alu and facilitate detection of its transcript binding with the lasso generated by transcription of the gene transcription framework (including the inserted sequence). In later experiments to verify the mechanism of action, the wavy line represents the transcription terminator. The double underline represents a 6TTTTAA repeat used to convert RNA to DNA, a common repeating sequence in genomes. You can choose to add or not add multiple sequences; in this embodiment, we chose to add 6. The sequence was obtained by chemical synthesis and designated Alu2. Alu2, plasmid pSIL-eGFP-MMP2-1, and plasmid pSIL-eGFP-MMP2-2 were each digested with SalI. The digestion reaction system is shown in Table 13: Table 13. Digestion reaction system The reaction was incubated at 37°C for 3 hours, followed by a 10-minute incubation at 80°C to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested Alu2 was ligated with the plasmid vectors pSIL-eGFP-MMP2-1 and pSIL-eGFP-MMP2-2, and the reaction system is shown in Table 14: Table 14. Linkage reaction systems The reaction conditions were incubation at 16°C for 16 hours, followed by incubation at 70°C for 10 minutes to inactivate the ligase. The plasmids pSIL-eGFP-MMP2-1-Alu2 and pSIL-eGFP-MMP2-2-Alu2 were obtained by electrophoresis and purification. Plasmids were sequenced to confirm their integrity. The pSIL-eGFP plasmid itself contains the U6 promoter, which is an RNA polymerase III-dependent promoter. Therefore, if an Alu sequence is inserted after the U6 promoter, transcription of the Alu sequence (Alu element) may be initiated by RNA polymerase III. pSIL-eGFP-MMP2-1-Alu2 or pSIL-eGFP-MMP2-2-Alu2 was transfected into U251 (human glioma) cells to test the insertion efficiency of the randomly designed exogenous sequences. To improve the insertion efficiency, pBS-L1PA1-CH-mneo, a plasmid expressing ORF1p and ORF2p (LINE), was co-transfected into U251 cells, and a corresponding control group was designed. The co-transfected plasmids for the experimental and control groups are listed in Table 15. Table 15 Grouping As shown in the groupings, control group 1 consisted of co-transfected pSIL-eGFP and pBS-L1PA1-CH-mneo, which lacked the gene transcription framework sequence and Alu2 sequence. Experimental group 5 consisted of co-transfected pSIL-eGFP-MMP2-1-Alu2 and pBS-L1PA1-CH-mneo, which contained a gene transcription framework with long sequences upstream and downstream of the target site in the MMP2 gene, an Alu2 sequence, and pBS-L1PA1-CH-mneo. Experimental group 6 consisted of co-transfected pSIL-eGFP-MMP2-1 and pBS-L1PA1-CH-mneo, which contained a gene transcription framework with long sequences upstream and downstream of the target site in the MMP2 gene, and pBS-L1PA1-CH-mneo, which lacked the Alu2 sequence. In experimental group 7, pSIL-eGFP-MMP2-2-Alu2 and pBS-L1PA1-CH-mneo were co-transfected, containing a gene transcription framework with a short sequence upstream and downstream of the target site in the MMP2 gene, an Alu2 sequence, and pBS-L1PA1-CH-mneo. In experimental group 8, pSIL-eGFP-MMP2-1-Alu2 was transfected, but pBS-L1PA1-CH-mneo was not. This gene transcription framework contains a long sequence upstream and downstream of the target site in the MMP2 gene, and an Alu2 sequence, but pBS-L1PA1-CH-mneo did not. Each group contained three parallel 6-well plates cultured with U251 cells. For the transfection procedure, U251 cells were passaged and spread into 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each plate of cells, 48 ​​μg or 96 μg of the constructed plasmid was diluted in 300 μL of serum-free DMEM and mixed thoroughly (48 μg if only one plasmid was transfected; 48 μg if two plasmids were co-transfected, for a total of 96 μg of the two plasmids, depending on the experimental group). At the same time, 120 μL of Entranster-H4000 reagent was diluted in 300 μL of serum-free DMEM, mixed thoroughly, and allowed to stand at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and allowed to stand at room temperature for 15 minutes to create the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM culture medium containing 10% fetal bovine serum per well for transfection. When the cells are about 90% confluent, the above procedure is repeated after passage until the cells are about 90% confluent and then removed for the subsequent procedure. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for detecting the GAPDH gene is shown in SEQ ID NO: 7 as follows: 5'-CACTGCCACCCAGAAGACTG-3'. The downstream primer sequence is shown in SEQ ID NO: 8 as follows: 5'-CCTGCTTCACCACCTTCTTG-3'. Primer pair 4 was designed, and the upstream primer sequence of primer pair 4 is shown as SEQ ID NO: 18: 5'-TTTCAGGGTCTAGGTGGC-3'; the downstream primer sequence is shown as SEQ ID NO: 19: 5'-AAATGCTTTCTCCGCTCT-3'. The upstream primer sequence of primer pair 4 is located in the complete MMP2 gene, and the upstream sequence of the insertion site (target site) used on the plasmid is located only in the genome, not within the plasmid, and the downstream primer sequence of primer pair 4 is located in the randomly designed non-homologous sequence to be inserted (the inserted sequence). The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 16. Table 16 qPCR reaction system JPEG2024504592000019.jpg80107 Cell DNA templates were extracted from the control group 1 and experimental groups 5-8 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 4: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 50°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. The exponential growth phase was observed in the amplification curves for the detection of insertion of GAPDH and the inserted sequence, and it was confirmed that they were nearly parallel. -ΔΔCt The results are shown in Table 17. The PCR products were confirmed to be correct by sequencing. Table 17 Results of primer pair 4 (n=3, x±s) Compared with other groups (N / A calculated according to 40.00), the relative copy number in experimental group 5 was significantly higher than the other groups, with statistical significance (P < 0.05). This suggests that gene editing is most efficient due to longer upstream or downstream sequences of the insertion site (target site) and appropriate expression of Alu elements (SINEs) and / or ORF1p and / or ORF2p (LINEs) in the gene transcription framework. The relative copy number amounts in experimental groups 6, 7, and 8 were higher than control group 1 (N / A calculated according to 40.00). All results were statistically significant (P < 0.05), suggesting that gene editing is still effective but less efficient. This is possible under the following conditions: the upstream or downstream sequence of the insertion site (target site) in the gene transcription framework is short, the cell itself has low or no expression of Alu elements (SINEs), and ORF1p and ORF2p (LINEs). Sufficient sequences on either side of the insertion site (target site) in the gene transcription framework are required to facilitate lariat formation to ensure efficient insertion. Editing efficiency can be improved by lengthening the upstream and / or downstream sequence of the target site or by additionally expressing ORF1p, ORF2p (LINEs), and / or Alu elements (SINEs). Example 4: Detecting connections between a lariat structure formed by a gene transcription framework (containing an inserted exogenous sequence) and an RNA fragment containing a SINE subsequence (e.g., an Alu element) (transcription of an Alu element cleaved at a natural shearing site) Extraction of total RNA from transfected cells in Experimental Group 5 and Control Group 1 in Example 3: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Transfer the cell-containing solution to an RNase-free centrifuge tube and centrifuge at 300 g for 5 minutes. Collect the pellet and aspirate all of the supernatant. Perform total RNA extraction according to the instructions of the magnetic bead method tissue / cell / blood total RNA extraction kit. Reverse transcription synthesis of cDNA template: cDNA is synthesized according to the instructions of the FastKing cDNA First Strand Synthesis Kit, and the concentration of the synthesized cDNA is determined by UV spectrophotometer for subsequent testing. qPCR testing: The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 5 was designed to detect the junction between the transcription lariat structure containing the exogenous sequence to be inserted and the RNA fragment containing the partial Alu sequence produced by transcription of the Alu element. The upstream primer sequence is shown in SEQ ID NO: 20: 5'-GGCATAATGATGTGGCTGTT-3'. The upstream primer is located on the exogenous sequence to be inserted, and the downstream primer sequence is shown in SEQ ID NO: 21: 5'-TCTGTTGGCTCGCTCTTG-3'. The downstream primer is located in the non-homologous sequence (18 bp) after the Alu sequence constructed in the plasmid. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 18. Table 18 qPCR reaction system JPEG2024504592000022.jpg80107 Reactions were prepared on ice. After preparation, the reaction tubes were capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 52°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. The exponential growth cycles in the amplification curves of GAPDH and the binding between the lariat structure containing the inserted sequence and the RNA fragment containing the partial Alu were observed and confirmed to be nearly parallel. -ΔΔCt The results are shown in Table 19. The PCR products were confirmed to be correct by sequencing. Table 19 Results of primer pair 5 (n=3, x±s) Compared with control group 1 (N / A calculated according to 40.00), the relative copy number in experimental group 5 was significantly higher with statistical significance (P < 0.05), indicating that the lariat construct containing the inserted exogenous sequence was indeed linked to the transcript of the Alu sequence (i.e., the RNA fragment containing the partial Alu). Table 19 shows that the transcript of the Alu sequence (i.e., the RNA fragment containing the partial Alu) was indeed linked to the transcript containing the inserted sequence. Example 5 tests the targeting accuracy of the gene editing technology of the present invention The sequence shown in SEQ ID NO: 15 is replaced with "CGATGA" for 5 to 10 bp (a total of 6 bp sequence) upstream of the randomly designed non-homologous sequence (i.e., the sequence to be inserted) to obtain the sequence shown in SEQ ID NO: 22: CTAGCTAGCTAGAGCATGGCGATGGATACCCCTTTGACGGTAAGGACGGACTCCTGGCTCATGCCTTCGCCCCAGGCACTGGTGTTGGGGGAGACTCCCATTTTGATGACGATGAGCTATGGACCTTGGGAGAAGGCCAAGGTGAGAAAGGGGCCCTCTGCATGCCCCAGACCTTCTCTCCTGTCCTCTCTCCACTCCATTTGCTTG CGATGA AGAG CCTGTGGGCCTTGCTCAGAGCGGAGAAAGCATGGCATAATGATGTGGCTGTTTTGTTTGTACAAGATCCGCAGACGTGTAAATGTTCCTGCAAAAACACAGACThe original wavy portion of this sequence, GACCAG, was replaced with CGATGA, and the remaining sequence was identical to Seq ID No. 15. This sequence was obtained by chemical synthesis and designated MMP2-3. MMP2-3 was prepared using the method of Example 3 to obtain the plasmid pSIL-eGFP-MMP2-3-Alu2. The plasmid pSIL-eGFP-MMP2-3-Alu2 was transfected into U251 cells (human glioma), and the plasmid pBS-L1PA1-CH-mneo (LINE) expressing ORF1p and ORF2p was also co-transfected into U251 cells as an experimental group. The aforementioned cells co-transfected with pSIL-eGFP-MMP2-1-Alu2 and pBS-L1PA1-CH-mneo served as a control group. The specific groupings are shown in Table 20. Table 20 Grouping Each group contained three parallel plates, each containing U251 cells cultured in a 6-well plate. The transfection method and post-transfection DNA extraction were the same as in Example 3. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. PCR detection using primer pair 4, and qPCR detection using the qPCR reaction system and reaction cycle of Example 3. The exponential growth phase was observed in the amplification curves for the detection of insertion of GAPDH and the inserted sequence, and it was confirmed that they were nearly parallel. -ΔΔCt The results are shown in Table 21. The PCR products were confirmed to be correct by sequencing. Table 21 Results of primer pair 4 (n=3, x±s) JPEG2024504592000025.jpg147169As can be seen from Table 21, the relative copy number of the experimental group was significantly lower than that of the control group (N / A calculated according to 40.00), with statistical significance (P<0.05). This indicates that if the upstream sequence of the insertion site (target site) on the vector does not match the upstream sequence of the insertion site (target site) on the genome, it is difficult to insert the sequence into the target site on the genome. Therefore, the gene editing performed by the present invention has high targeting accuracy. Example 6 Application of the SINE sequence (Alu sequence as an example) direct ligation method to insert an exogenous sequence to be inserted The IT15 gene is the causative gene for Huntington's disease, and in this example, an exogenous sequence is inserted into the IT15 gene to confirm the genomic sequence insertion technique "direct ligation of SINE sequences via DNA (using Alu sequences as an example)." A 160 bp sequence of the gene IT15 in the human genome was selected as shown in SEQ ID NO: 23: ATGCTATTCATAATCACATTCGTTTGTTTGAACCTCTTGTTATAAAAGCTTTAAAACAGTACACGACTACAACATGTGTGCAGTTACA G*A AGCAGGTTTTAGATTTGCTGGCGCAGCTGGTTCAGTTACGGGTTAATTACTGTCTTCTGGATTCAGATCA, where * is the selected insertion site (target site), the upstream sequence of the insertion site of the IT15 gene before the insertion site (upstream sequence of the target site), and the downstream sequence of the insertion site of the IT15 gene after the insertion site (downstream sequence of the target site). A randomly designed non-homologous sequence was added to the insertion site as the inserted sequence, and a partial Alu sequence was ligated downstream of the target site to form a gene transcription framework. To enable the gene transcription framework to be inserted into an expression vector, restriction enzyme NheI digestion sites and protective bases were added to both ends. The complete sequence is shown in SEQ ID NO: 24. CTAGCTAGCTAGATGCTATTCATAATCACATTCGTTTGTTTGAACCTCTTGTTATAAAAGCTTTAAAACAGTACACGACTACAACATGTGTGCAGTTACAG GAAGTTCATGGATGTCTATCAGCGCAGCTACTGCCATCCAATCGAGACCCTGGTGGACAT AAGCAGGTTTTAGATTTGCTGGCGCAGCTGGTTCAGTTACGGGTTAATTACTGTCTTCTGGATTCAGATCAACTAAAAATACAAAAAATTAGCCGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCACCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATCCTAGCTAGCTAG, where the underscore indicates a randomly designed exogenous non-homologous sequence (i.e., the sequence to be inserted). The sequence is 60 bp long, with NheI digestion sites and protective bases (in bold italics) at both ends, and a partial Alu sequence (hatched) between the downstream sequence of the IT15 gene insertion site (downstream sequence of the target site) and the 3′ NheI digestion site and protective base sequence. The sequence was obtained by chemical synthesis and designated IT15-1. The choice of attaching a partial Alu element downstream of the target site was chosen to mimic a specific state formed by a specific structure in an organism: a structure retaining reverse transcription function, generated by intracellular action (cleavage of the SINE transcript at its natural shearing site), coupled with a lariat structure generated by pre-mRNA shearing. As shown in Figure 17, IT15-1 was inserted into the plasmid vector pBS-L1PA1-CH-mneo to construct the plasmid pBS-L1PA1-CH-mneo-IT15-1. The specific process is as follows: IT15-1 and pBS-L1PA1-CH-mneo were digested separately, and the reaction system is shown in Table 22: Table 22 Digestion reaction system The reaction conditions were incubation at 37°C for 1 hour, followed by incubation at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested IT15-1 was ligated with the plasmid vector pBS-L1PA1-CH-mneo, and the reaction system is shown in Table 23: Table 23 Linkage reaction systems The reaction was incubated at 16 °C for 16 hours, followed by 10 minutes at 70 °C to inactivate the ligase. The resulting plasmid, pBS-L1PA1-CH-mneo-IT15-1, was electrophoresed and purified. The plasmid was sequenced to confirm its integrity. The pBS-L1PA1-CH-mneo plasmid has its own CMV promoter, so transcription can be initiated by RNA polymerase II as long as the expression framework is inserted into the CMV promoter. Experimental groups: the group transfected with pBS-L1PA1-CH-mneo-IT15-1 plasmid was designated as experimental group 10; the group transfected with unmodified pBS-L1PA1-CH-mneo plasmid was designated as control group 3. Each group contained three parallel 6-well plates cultured with HeLa cells. For the transfection procedure, HeLa cells were passaged and spread onto 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each 6-well plate, 48 μg of the constructed plasmid was diluted with 300 μL of serum-free DMEM and mixed thoroughly. At the same time, 120 μL of Entranster-H4000 reagent was diluted with 300 μL of serum-free DMEM, mixed thoroughly, and allowed to stand at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and allowed to stand at room temperature for 15 minutes to form the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM medium containing 10% fetal bovine serum per well for transfection. When the cells reached approximately 90% confluence, the above procedure was repeated after passage. Once the cells reached approximately 90% confluence, they were removed for subsequent procedures. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 6: The upstream primer sequence is shown in SEQ ID NO: 25: 5'-GAAATTGGTTTGAGCAGGAG-3'; the downstream primer sequence is shown in SEQ ID NO: 26: 5'-CGATTGGATGGCAGTAGC-3'. The upstream primer sequence of primer pair 6 is located in the intact IT15 gene, and furthermore, the upstream sequence of the upstream insertion site (target site) used on the plasmid is located only in the genome, not in the plasmid, while the downstream primer sequence of primer pair 6 is located in a randomly designed non-homologous sequence (the inserted sequence). The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 24. Table 24 qPCR reaction system JPEG2024504592000028.jpg89118 Cell DNA templates were extracted from the control group 3 and experimental group 10 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 6: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 50°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. The exponential growth phase was observed in the amplification curves for the detection of insertion of GAPDH and the inserted sequence, and it was confirmed that they were nearly parallel. -ΔΔCt The results are shown in Table 25. The PCR products were confirmed to be correct by sequencing. Table 25 Results of primer pair 6 (n=3, x±s) JPEG2024504592000029.jpg89169JPEG2024504592000030.jpg60169Table 25 shows that the relative copy number of experimental group 10 is significantly higher than that of control group 3 (N / A is calculated according to 40.00), which is statistically significant (P<0.05), indicating that the inserted sequence is efficiently inserted into the target site on the genome. Example 7 tests the targeting accuracy of the gene editing technology of the present invention The sequence shown in SEQ ID NO: 27 is obtained by replacing 10 bp to 15 bp (a total of 6 bp sequence) upstream of the randomly designed non-homologous sequence (i.e., the sequence to be inserted) in the sequence shown in SEQ ID NO: 24 with "GGACAT". CTAGCTAGCTAGATGCTATTCATAATCACATTCGTTTGTTTGAACCTCTTGTTATAAAAGCTTTAAAACAGTACACGACTACAACA GGACAT CAGTTACAG GAAGTTCATGGATGTCTATCAGCGCAGCTACTGCCATCCAATCGAGACCCTGGTGGACAT The sequence was: AAGCAGGTTTTAGATTTGCTGGCGCAGCTGGTTCAGTTACGGGTTAATTACTGTCTTCTGGATTCAGATCAACTAAAAATACAAAAAATTAGCCGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCACCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATCCTAGCTAGCTAG. The original wavy portion, TGTGTG, was replaced with GGACAT, and the remaining sequence was identical to Seq ID No. 24. This sequence was obtained by chemical synthesis and designated IT15-2. IT15-2 was inserted into the plasmid vector pBS-L1PA1-CH-mneo to construct the plasmid pBS-L1PA1-CH-mneo-IT15-2, as described in Example 6. The Hela cell group transfected with pBS-L1PA1-CH-mneo-IT15-2 plasmid was designated as experimental group 11; the Hela cell group transfected with pBS-L1PA1-CH-mneo-IT15-1 plasmid was designated as control group 4. Each group contained three parallel 6-well plates containing Hela cells. The method of Example 6 was used for transfection and extraction of transfected cell DNA for qPCR detection. The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 6 was used for qPCR, and the reaction system and reaction cycle were the same as those in Example 6. An exponential growth phase was observed in the amplification curves for detecting the insertion of GAPDH and the inserted sequence, and it was confirmed that they were nearly parallel. -ΔΔCt The results are shown in Table 26. The PCR products were confirmed to be correct by sequencing. Table 26 Results of primer pair 6 (n=3, x±s) The relative copy number in the experimental group was significantly lower than that in the control group (N / A calculated according to 40.00), and this was statistically significant (P<0.05). This indicates that it is difficult to insert a sequence into the target site on the genome when the upstream sequence of the insertion site (target site) on the vector does not match the upstream sequence of the insertion site (target site) on the genome. Conclusion: The genomic sequence insertion performed by the present invention has high targeting accuracy. Examples 1 to 6 demonstrate that the DNA-mediated insertion of exogenous sequences into specific genomic sites can effectively edit eukaryotic cells (such as cell lines and primary cells) and target the inserted sequences to the target site with high efficiency and precision. The feasibility of editing cells from different tissues demonstrates that this method can be applied to a variety of cells, tissues, and organisms (organisms). Example 8 DNA-mediated deletion (deletion) of a genomic sequence (sequence to be deleted) The sequence of the gene MINK1 in the human genome is randomly selected as shown in SEQ ID NO: 28: AGAGAATGAGGGGCCCCTTTTTCTCTCTGGTGGCTCAGGCCCAACTCCCTTCCTACTGGGGAGGCTCACTCCCTCCCCTTTCCCCTCTCCCCCTGGAATGCCCTGCCTCCTGCTGAAAATCCCTCAGGAAGCTCTTCACCTGTCACCTGTTACGGCCAGGTGCTCTGCAGGTTGCTCTG GGGAGATGGGATCTGATGGCCCTCCTGCCTGGGATGCTGTCCGTGATCCTTTTACCTGGGTTTTTCTCTAAGATGCTGGAAGATGGAATCGGGTTCTTCAGGATGGTGGTGGGGTAAAGGAGGGTGCTGGGGTGTCTGGGTCGGGCCAGGACCACAGCTGGCTCAGGCAAGTCCTGTGTGTGCACGCAGGGATGTGAGGCAAGGGAGCAGAGGTGACTCCCCACACTGACCCCTCCCTCTGTGTCTTCACAGTGGATTTTGGGGTGAGTGCTCAGCTGGACCGCACCGTGGGCAGACGGAACACTTTCATTGGGACTCCCTACTGGATGGCTCCAGAGGTCATCGCCTGTGATGAGAACCCTGATGCCACCTATGATTACAGGGTATGGAGTGGAAAGTTGGGAGCATGGGGGCTGCCAAGGGCGGGAAGCAATATGGGGACCACGGGGCCTGAGCAGGCTGGGGAACAGAGGAAGGTCAGATGATGTTAGCAGTGAGGGGCTGGGGAACATCTTACGGCAAGGCAAGTGTGGGTGGGAAGATGGGATGGGTTGGAAGGCACTGCTGCAGGAATGGGTGTGGCCCAGGAAGGCTCCTGAGAGGCCAGGATGGTGGGTGAAGAGAGGTTGCAGGGCAGAGTTGTCAGGAATATTCACTTGTTCCTTCTTTCCCGTCTATAGAGTGATATTTGGTCTCTAGGAATCACAGCCATCGAGATGGCAGAGGGAGCCCCCCGTAAGTTCTGAGTCTGCC GGGAGTGGGAGGGGAGGGAAAGGAAGGGCCCAGAGAGTGGCTGTAGGGAGGAGGTGGGTCCTGGGACCCTGCCGAGGAAGGGTCCTGTAGCTCCCAGTGCAGTGAAAGGGACTGAGGGTGTCTCCTCTGTGTCCAGCTCTGTGTGACATGCACCCCATGCGAGCCCTCTTCCTCATTCCTCGGAACCCTCCGCCCAGGCTCAAGTCCAAGAAGTG, where the underlined portion is the sequence to be deleted in the genome, the upstream sequence immediately adjacent to the 5' end of the sequence to be deleted in the genome before the deleted sequence, the downstream sequence immediately adjacent to the 3' end of the sequence to be deleted in the genome after the deleted sequence, and the shaded portion is the 3' sequence of the sequence to be deleted in the genome. The sequence was constructed in the order of 3' sequence of the sequence to be deleted + upstream sequence immediately adjacent to the 5' end of the sequence to be deleted + downstream sequence immediately adjacent to the 3' end of the sequence to be deleted, with NheI digestion sites and corresponding protecting bases added to both ends, and the sequence is shown in SEQ ID NO: 29: CTAGCTAGCTAGAGTGATATTTGGTCTCTAGGAATCACAGCCATCGAGATGGCAGAGGGAGCCCCCCGTAAGTTCTGAGTCTGCCAGAGAATGAGGGGCCCCTTTTTCTCTCTGGTGGCTCAGGCCCAACT CCCTTCCTACTGGGGAGGCTCACTCCCTCCCCTTTCCCCTCTCCCCCTGGAATGCCCTGCCTCCTGCTGAAAATCCCTCAGGAAGCTCTTCACCTGTCACCTGTTACGGGCCAGGTGCTCTGCAGGTTGCTC The sequence was synthesized by chemical synthesis and designated MINK1-1. At the same time, the upstream sequence immediately adjacent to the 5' end of the sequence to be deleted in SEQ ID NO: 29 is replaced with a sequence that is not homologous to the MINK1 gene, resulting in the sequence shown in SEQ ID NO: 30: CTAGCTAGCTAGAGTGATATTTGGTCTCTAGGAATCACAGCCATCGAGATGGCAGAGGGAGCCCCCCGTAAGTTCTGAGTCTGCC CATTGTTATAATGATTAAACAAGTTATATATGAAAAGATTAAAACAGTGTTGCTCCATAATAAATGCTGTTTTTACTGTGATTATTATTGTTGTTATCCCTATCATTATCATCACCATCTTAACCCTTCCCTGTTTTGCTCTTTTCTCTCTCCCTACCCATTGCAGACCAAAGAAAGATAThe sequence was synthesized chemically and designated MINK1-2. MINK1-1 and MINK1-2 were each inserted into the plasmid vector pSIL-eGFP to construct the plasmids pSIL-eGFP-MINK1-1 and pSIL-eGFP-MINK1-2 as follows. MINK1-1, MINK1-2, and the plasmid pSIL-eGFP were digested separately, and the reaction system is shown in Table 27: Table 27. Digestion reaction system The reaction conditions were incubation at 37°C for 1 hour, followed by incubation at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested MINK1-1 or MINK1-2 was ligated with the plasmid vector pSIL-eGFP, and the reaction system is shown in Table 28: Table 28 Linkage reaction systems The reaction conditions were incubation at 16 °C for 16 hours, followed by incubation at 70 °C for 10 minutes to inactivate the ligase. The plasmids pSIL-eGFP-MINK1-1 and pSIL-eGFP-MINK1-2 were obtained by electrophoresis and purification. Plasmids were sequenced to confirm their authenticity. The Alu1 prepared in Example 1, the plasmid pSIL-eGFP-MINK1-1, and the plasmid pSIL-eGFP-MINK1-2 were digested with SalI and ligated. The reaction system and conditions were the same as in Example 1 to obtain pSIL-eGFP-MINK1-1-Alu1 and pSIL-eGFP-MINK1-2-Alu1. The effectiveness of deleting the target sequence was tested by transfecting HeLa cells with pSIL-eGFP-MINK1-1-Alu1 or pSIL-eGFP-MINK1-2-Alu1. To improve the deletion efficiency, the plasmid pBS-L1PA1-CH-mneo, which expresses ORF1p and ORF2p (LINE), was co-transfected into HeLa cells, and corresponding control groups were designed. The group transfected with pSIL-eGFP-MINK1-1-Alu1 and pBS-L1PA1-CH-mneo was designated experimental group 12, and the group transfected with pSIL-eGFP-MINK1-2-Alu1 and pBS-L1PA1-CH-mneo was designated control group 5. Each group contained three parallel 6-well plates containing HeLa cells. Plasmid transfection is carried out according to the method of Example 1, and transfected cell DNA is extracted. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 7: The upstream primer sequence is shown in SEQ ID NO: 31: 5'-ACAGGGTATGGAGTGGAAAG-3'; the downstream primer sequence is shown in SEQ ID NO: 32: 5'-ATAGACGGGAAAGAAGGAAC-3'. The upstream primer of primer pair 7 is located in the sequence of the MINK1 gene that will soon be deleted on the genome and is not present in the plasmid, and the downstream primer is located in the sequence of the MINK1 gene that will soon be deleted on the genome and is not present in the plasmid. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 29. Table 29 qPCR reaction system JPEG2024504592000034.jpg80107 Cell DNA templates were extracted from the control group 5 and experimental group 12 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 7: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 50°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. After observing the exponential growth cycles of the amplification curves of GAPDH and the sequence to be deleted and confirming that they are nearly parallel, the data obtained are analyzed by 2 -ΔΔCt The results are shown in Table 30. The PCR products were confirmed to be correct by sequencing. Table 30 Results of primer pair 7 (n=3, x±s) JPEG2024504592000035.jpg74169JPEG2024504592000036.jpg68169The relative copy number (N / A calculated at 40.00) of experimental group 12 was significantly lower than that of control group 5, and the experimental results were statistically significant (P<0.05), indicating that the deleted sequence on the genome was deleted. Example 9: Deleting a SINE sequence (using an Alu sequence as an example) using the direct linking method The FMR1 gene is associated with inherited mental retardation - Fragile X syndrome, and one of the sequences was selected, as shown in SEQ ID NO: 33: ACTTGCTGAGTACCCAAGGAAAGTGTGCTTGTATTTATGGGCGTCTATTTTCAGAGCACTAATTATTGCTGAATTAGAACAGAAATATAGGAAAACTGATTTTTACAAGGAGCTTCAAAGCAATCTCAGGTAGTTTCTGATTATGTATCTCTGCCTACCTCGGGGTACATAGACAGGGTTACAATTTGGTTGAGGATATATGACATGTGGTTTTTTAAAGACACCTAGGGGCATTTTAAGAAAATTTCCTCGATATCTGAAAATCTGTAGATTTCAAAATTATGTTAATCATGAAATATTCTGTGTTGTAATTTTTGTGTAGGTGTATTCCAGAGCAAATGAAAAAGAGCCTTGCTGTTGGTGGTTAGCTAAAGTGAGGATGATAAAGGGTGAGGTAGGAAAATGCCTATTTAAATTTTTTTCTTATATT GTTTCCTTTTTTTAAACCCAGGTTGTACATTCCCGTGTGGGATTTCTATTTTGAAGTAATATCTAATTTTGAGTAATTTAATTAAAATGTTTTCACTATGTGTTCAGTATGTTTCTGTTGGTCATAAATTTTTTCACATAGATTATTTATTTTAAAATAACTGAATAGGGAGAACTTCTTATTCTTACTTTAAAAATTGTGATTAGAAGTGACTTTTATTTATTTCTCAGTTTTATGTGATAGAATATGCAGCATGTGATGCAACTTACAATGAAATTGTCACAATTGAACGTCTAAGATCTGTTAATCCCAACAAACCTGCCACAAAAG ATACTTTCCATAAGATCAAGCTGGATGTGCCAGAAGACTTACGGCAAA, where the underlined portion is the sequence to be deleted in the genome, the upstream sequence immediately adjacent to the 5' end of the sequence to be deleted in the genome before the sequence to be deleted, the downstream sequence immediately adjacent to the 3' end of the sequence to be deleted in the genome after the sequence to be deleted, and the shaded portion is the 3' sequence of the sequence to be deleted in the genome. The sequence is assembled in the following order: 3' sequence of the sequence to be deleted + upstream sequence immediately adjacent to the 5' end of the sequence to be deleted + downstream sequence immediately adjacent to the 3' end of the sequence to be deleted + partial Alu sequence, with NheI digestion sites and corresponding protecting bases added to both ends, and the sequence is shown in SEQ ID NO: 34: CTAGCTAGCTAGTTTATGTGATAGAATATGCAGCATGTGATGCAACTTACAATGAAATTGTCACAATTGAACGTCTAAGATCTGTTAATCCCAACAAACCTGCCACAAAAG ACTTGCTGAGTACCCAAGGAAAGTGTGCTTGTATTTATGGGCGTCTATTTTCAGAGCACTAATTATTGCTGAATTAGAACAGAATATAGGAAAACTGAT ATACTTTCCATAAGATCAAGCTGGATGTGCCAGAAGACTTACGGCAAAACTAAAAATACAAAAAATTAGCCGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCACCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATCCTAGCTAGCTAG. The italicized and bolded parts at both ends are NheI digestion sites and the corresponding protected bases. The shaded parts are partial Alu sequences. The sequence was obtained by chemical synthesis and designated FMR1-1. At the same time, the upstream sequence immediately adjacent to the 5' end of the sequence to be deleted in SEQ ID NO: 34 is replaced with a sequence that is not homologous to the FMR1 gene, resulting in the sequence shown in SEQ ID NO: 35: CTAGCTAGCTAGTTTATGTGATAGAATATGCAGCATGTGATGCAACTTACAATGAAATTGTCACAATTGAACGTCTAAGATCTGTTAATCCCAACAAACCTGCCACAAAAG AGGTTGCTCCATAATAAATGCTGTTTTTACTGTGATTATTATTGTTGTTATCCCTATCATTATCATCACCATCTTAACCCTTCCCTGTTTTGCTCTTTT ATACTTTCCATAAGATCAAGCTGGATGTGCCAGAAGACTTACGGCAAAACTAAAAATACAAAAAATTAGCCGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCACCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATCCTAGCTAGCTAG, where the underscore represents a replacement sequence that is not homologous to the FMR1 gene. The sequence was obtained by chemical synthesis and designated FMR1-2. FMR1-1 and FMR1-2 were each inserted into the plasmid vector pBS-L1PA1-CH-mneo to construct the plasmids pBS-L1PA1-CH-mneo-FMR1-1 and pBS-L1PA1-CH-mneo-FMR1-2 as follows. FMR1-1, FMR1-2, and the plasmid pBS-L1PA1-CH-mneo were digested separately, and the reaction system is shown in Table 31: Table 31 Digestion reaction system The reaction was incubated at 37°C for 1 hour, then at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested FMR1-1 or FMR1-2 was ligated with the plasmid vector pBS-L1PA1-CH-mneo, and the reaction system is shown in Table 32: Table 32 Linkage reaction systems The reaction conditions were incubation at 16°C for 16 hours, followed by incubation at 70°C for 10 minutes to inactivate the ligase. The plasmids pBS-L1PA1-CH-mneo-FMR1-1 and pBS-L1PA1-CH-mneo-FMR1-2 were obtained by electrophoresis and purification. Plasmids were sequenced to confirm their authenticity. The group transfected with pBS-L1PA1-CH-mneo-FMR1-1 was designated experimental group 13, and the group transfected with pBS-L1PA1-CH-mneo-FMR1-2 was designated control group 6. Each group contained three parallel plates, each consisting of a 6-well plate cultured with HeLa cells. For the transfection procedure, HeLa cells were passaged and spread onto 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each 6-well plate, 48 μg of the constructed plasmid was diluted with 300 μL of serum-free DMEM and mixed thoroughly. At the same time, 120 μL of Entranster-H4000 reagent was diluted with 300 μL of serum-free DMEM, mixed thoroughly, and allowed to stand at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and allowed to stand at room temperature for 15 minutes to form the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM medium containing 10% fetal bovine serum per well for transfection. When the cells reached approximately 90% confluence, the above procedure was repeated after passage. Once the cells reached approximately 90% confluence, they were removed for subsequent procedures. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 8: The upstream primer sequence is shown in SEQ ID NO: 36: 5'-ACAGGGTTACAATTTGGT-3'; the downstream primer sequence is shown in SEQ ID NO: 37: 5'-CATTTGCTCTGGAATACAC-3'. The upstream primer of primer pair 8 is located on the sequence of the FMR1 gene that will soon be deleted on the genome and is not present on the plasmid, and the downstream primer is located on the sequence of the FMR1 gene that will soon be deleted on the genome and is not present on the plasmid. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 33. Table 33 qPCR reaction system JPEG2024504592000039.jpg80108 Cell DNA templates were extracted from the control group 6 and experimental group 13 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 8: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 45°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. After observing the exponential growth cycles of the amplification curves of GAPDH and the sequence to be deleted and confirming that they are nearly parallel, the data obtained are analyzed by 2 -ΔΔCt The results are shown in Table 34. The PCR products were confirmed to be correct by sequencing. Table 34 Results of primer pair 8 (n=3, x±s) JPEG2024504592000040.jpg140169The relative copy number (N / A calculated at 40.00) of experimental group 13 was significantly lower than that of control group 6, and the experimental results were statistically significant (P<0.05), indicating that the deleted sequence on the genome was deleted. From the above embodiment, it can be seen that the present invention can be used to delete any sequence in any region of the genome. At the same time, if sequencing obtains the CNV ends of each gene (i.e., if the corresponding gene sequence connects a portion of the SINE sequence), the CNV ends can also be edited: CNVs can be edited by insertion or deletion (CNV ends), and the resulting expression changes can change the state of cells, tissues, or organisms. Example 10 Insertion of foreign sequences inserted into the genome via RNA The sequence of gene IT15 in the human genome was selected, and the corresponding sequence was designed in the following order: NheI digestion recognition site and protecting bases + upstream sequence of insertion site (target site) + sequence to be inserted + downstream sequence of insertion site (target site) + partial Alu sequence + NheI digestion recognition site and protecting bases, as shown in SEQ ID NO: 38: CTAGCTAGCTAGATGCTATTCATAATCACATTCGTTTGTTTGAACCTCTTGTTATAAAAGCTTTAAAACAGTACACGACTAC GAAGTTCATGGATGTCTATCAGCGCAGCTACTGCCATCCAATCGAGAGCCCTGGTGGACATAACATGTGTGCAGTTACAGAAGCAGGTTTTAGATTTGCTGGCGCAGCTGGTTCAGTTACGGGTTAATTACTGTCTTCTGGATTCAGATCAACTAAAAATACAAAAAATTAGCCGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCACGCCGCTGCACTCCACCCTGGGCGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGATTAATAACTGCTGGAGATCCTAGCTAGCTAG, where the underscore indicates a randomly designed sequence (not homologous to the IT15 gene) to be inserted. The sequence is 60 bp long, with NheI digestion sites and protective bases (in bold italics) at both ends, and a partial Alu sequence (hatched) between the downstream sequence of the IT15 gene insertion site (target site) and the 3' NheI digestion site and protective base sequence. The sequence was obtained by chemical synthesis and designated IT15-3. IT15-3 was inserted into the plasmid vector pBS-L1PA1-CH-mneo to construct the plasmid pBS-L1PA1-CH-mneo-IT15-3 as follows. IT15-3 and plasmid pBS-L1PA1-CH-mneo were digested separately, and the reaction system is shown in Table 35: Table 35. Digestion reaction system The reaction was incubated at 37°C for 1 hour, then at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested IT15-3 was ligated with the plasmid vector pBS-L1PA1-CH-mneo, and the reaction system is shown in Table 36: Table 36 Linkage reaction systems The reaction was incubated at 16 °C for 16 hours, followed by 10 minutes at 70 °C to inactivate the ligase. The resulting plasmid, pBS-L1PA1-CH-mneo-IT15-3, was electrophoresed and purified. The plasmid was sequenced to confirm its integrity. Then, the pBS-L1PA1-CH-mneo-IT15-3 and pBS-L1PA1-CH-mneo plasmids were transfected into Hela cells, respectively. For the transfection procedure, HeLa cells were passaged and spread onto 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each 6-well plate, 48 μg of the constructed plasmid was diluted with 300 μL of serum-free DMEM and mixed thoroughly. At the same time, 120 μL of Entranster-H4000 reagent was diluted with 300 μL of serum-free DMEM, mixed thoroughly, and allowed to stand at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and allowed to stand at room temperature for 15 minutes to form the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM medium containing 10% fetal bovine serum per well for transfection. When the cells reached approximately 90% confluence, the above procedure was repeated after passage. Once the cells reached approximately 90% confluence, they were removed for subsequent procedures. Extraction of transfected cell RNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Transfer the cell-containing solution to an RNase-free centrifuge tube and centrifuge at 300g for 5 minutes. Collect the pellet and aspirate all the supernatant. Perform total RNA extraction according to the instructions of the magnetic bead method tissue / cell / blood total RNA extraction kit. Extraction of mRNA from total RNA: The concentration of the previously extracted total RNA was detected by UV spectrophotometer, 1000 ng of total RNA was diluted to 50 µL with nuclease-free ddHO, mRNA was extracted according to the instructions of the TIANSeq mRNA Capture Kit, and an aliquot of the sample was taken to detect the mRNA content (the above experimental procedure was repeated several times to obtain enough mRNA for transfection). Experimental groups: Experimental group 14 received mRNA extracted from Hela cells transfected with the pBS-L1PA1-CH-mneo-IT15-3 plasmid; control group 7 received mRNA extracted from Hela cells transfected with the pBS-L1PA1-CH-mneo plasmid. Each group contained three parallel 6-well plates containing Hela cells. The resulting mRNA was transferred into HeLa cells using Lipofectamine MessengerMAX transfection reagent, and 14 experimental and 7 control cells were cultured in 6-well plates. The first transfection was performed when the cells reached 40% confluence. For each well of transfected cells, 7.5 μL of Lipofectamine MessengerMAX transfection reagent was diluted with 125 μL of serum-free DMEM solution and incubated at room temperature for 10 minutes. 5 μg of the prepared mRNA was mixed with 125 μL of serum-free DMEM solution, followed by 125 μL of pre-diluted Lipofectamine MessengerMAX transfection reagent and incubation at room temperature for 5 minutes. The resulting mixture was added to each well of cultured cells and gently mixed. The above procedure was repeated when the cells reached 70% confluence. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 6: The upstream primer sequence is shown in SEQ ID NO: 25: 5'-GAAATTGGTTTGAGCAGGAG-3'; the downstream primer sequence is shown in SEQ ID NO: 26: 5'-CGATTGGATGGCAGTAGC-3'. The upstream primer sequence of primer pair 6 is located in the intact IT15 gene, and furthermore, the upstream sequence of the upstream insertion site (target site) used on the plasmid is located only in the genome, not in the plasmid, while the downstream primer sequence of primer pair 6 is located in a randomly designed non-homologous sequence (the inserted sequence). The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 37. Table 37 qPCR reaction system JPEG2024504592000043.jpg81107 Cell DNA templates were extracted from the control group 7 and experimental group 14 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 6: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 50°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. The exponential growth phase was observed in the amplification curves for the detection of insertion of GAPDH and the inserted sequence, and it was confirmed that they were nearly parallel. -ΔΔCt The results are shown in Table 38. The PCR products were confirmed to be correct by sequencing. Table 38 Results of primer pair 6 (n=3, x±s) JPEG2024504592000044.jpg67169JPEG2024504592000045.jpg83169Table 38 shows that the relative copy number of experimental group 14 was significantly higher than that of control group 7 (N / A calculated according to 40.00), which was statistically significant (P<0.05), indicating that the inserted sequence was efficiently inserted into the target site on the genome, indicating that gene editing using the RNA pathway of the present invention is effective. With the feasibility of complete RNA-mediated transcription, if the coding sequences of ORF2p and / or ORF1p are not added to the sequence to be imported into the editing system, an RNA product containing the upstream sequence of the insertion site (target site) + the sequence to be inserted + the downstream sequence of the insertion site (target site) + a SINE sequence (such as an Alu sequence), partial SINE sequence, or similar SINE sequence can bind to ORF2p in vitro or simultaneously bind to ORF1p and ORF2p and be transcribed into the cell (cytoplasm) by the RNP-mediated pathway. Example 11 CNV ends are fixed and edited according to the present invention Based on the widespread phenomenon of CNV expansion in living organisms, we can infer that each gene's CNV should have a terminal end. Specifically, a partial SINE (Alu) sequence is connected downstream of a corresponding gene segment, and a partial SINE (Alu) sequence (double-stranded DNA) is connected by different lassoes. This sequential insertion of gene sequences precedes the partial SINE sequence at the end of the CNV, gradually extending the CNV. Because exons must be excised from pre-mRNA, the end of an intron must form a lasso. Because the probability of generating a lasso that includes an exon that overlaps it is low, a relatively long period is required for genes with relatively low expression. The CNV terminus is located at the end of the gene's intron, bound to the partial SINE (Alu) sequence. In this example, the 3' sequence of an intron in the BRCA1 gene was randomly selected and selected as the sequence to be deleted in the following example. The 3' sequence of the BRCA1 gene intron is shown in SEQ ID NO: 39: CCCAGCTACTTGAGAGGCTGAGGCAGGGAGAATTGCTTGAACCAGGTAGGCGGAGGTTGCAGTGAGCCAAGATCGCACCACTGCACTCCAGCCTGGGGCAACAGAGCAAGACTGTCTCAAAAAAAATAAATAAATAAAATAAATTCTTAA ACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG , where the underlined portion is the upstream sequence immediately adjacent to the 5' end of the sequence to be deleted, the sequence after the underline is the sequence to be deleted (specifically, the 3' sequence of the intron of the BRCA1 gene), and the wavy line is the 3' terminal sequence of the sequence to be deleted (hence the name, deletion, as performed in the following embodiment). The sequence is constructed by adding the 3'-terminal sequence of the sequence to be deleted, a randomly designed sequence that is not homologous to the BRCA1 gene, and a partial Alu sequence, and then adding NheI digestion sites and protective bases to both ends of the sequence. Because natural shearing sites that cause Alu element transcripts to produce RNAs containing only partial Alu sequences have not been consistently reported in different literatures, to prevent insertion failure due to mismatches with partial Alu sequences at the CNV ends, three possible partial Alu sequences were synthesized and introduced (the difference is that their 5'-end sequences are different), and the relevant sequences were constructed as shown in SEQ ID NO:40, SEQ ID NO:41, and SEQ ID NO:42. The sequence SEQ ID NO: 40 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG GGGGGG The sequence is: AAAAATACAAAAAATTAGCCGGGCGTGATGGTGGGCGCCTGTAGTCCCAGCTACACGGGAGGCTGAGACAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCACCACTGCACTGCAGCCTGGGCGACAGAGCGAGACTCTGTCTCAAAAAAAAAAAAAAAGAAAGAAAAAAAACTAGCTAGCTAG, where the underscore indicates a randomly designed non-homologous sequence (sequence not homologous to the BRCA1 gene), NheI cleavage sites and protective bases (bold italics) at both ends of the sequence, and a partial Alu sequence, Alu3 (shaded area), between the non-homologous sequence and the 3'-terminal NheI cleavage site and protective base sequence. The sequence was obtained by chemical synthesis and designated BRCA1-1-Alu3. The sequence SEQ ID NO: 41 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG GGGGGG CTAAAAATACAAAAAATTAGCCGGGCGTGATGGTGGGCGCCTGTAGTCCCAGCTACACGGGAGGCTGAGACAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCACCACTGCACTGCAGCCTGGGCGACAGAGCGAGACTCTGTCTCAAAAAAAAAAAAAAAGAAAGAAAAAAAACTAGCTAGCTAG, where the underscore indicates a randomly designed non-homologous sequence (sequence not homologous to the BRCA1 gene), NheI cleavage sites and protective bases (bold italics) at both ends of the sequence, and a partial Alu sequence, Alu4 (shaded area), between the non-homologous sequence and the 3'-terminal NheI cleavage site and protective base sequence. The sequence was obtained by chemical synthesis and designated BRCA1-1-Alu4. The sequence SEQ ID NO: 42 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG GGGGGG The sequence is shown in bold and italicized text as follows: ACTAAAAATACAAAAAATTAGCCGGGCGTGATGGTGGGCGCCTGTAGTCCCAGCTACACGGGAGGCTGAGACAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCACCACTGCACTGCAGCCTGGGCGACAGAGCGAGACTCTGTCTCAAAAAAAAAAAAAAAGAAAGAAAAAAAACTAGCTAGCTAG, where the underscore indicates a randomly designed non-homologous sequence (sequence not homologous to the BRCA1 gene), NheI cleavage sites and protective bases at both ends of the sequence, and a partial Alu sequence, Alu5 (shaded area), between the non-homologous sequence and the 3'-terminal NheI cleavage site and protective base sequence. The sequence was obtained by chemical synthesis and designated BRCA1-1-Alu5. As shown in SEQ ID NO:43, SEQ ID NO:44 and SEQ ID NO:45, sequences that do not contain non-homologous sequences (sequences that are not homologous to the BRCA1 gene) are also designed. The sequence of SEQ ID NO: 43 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAGAAAAATACAAAAAATTAGCCGGGCGTGATGGTGGGCGCCTGTAGTCCCAGCTACACGGGAGGCTGAGACAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCACCACTGCACTGCAGCCTGGGCGACAGAGCGAGACTCTGTCTCAAAAAAAAAAAAGAAAGAAAAAAAACTAGCTAGCTAG, where SEQ ID NO: 43 is BRCA1-1-Alu3 without non-homologous sequences. The sequence was obtained by chemical synthesis and designated BRCA1-2-Alu3. SEQ ID NO: 44 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAGCTAAAAATACAAAAAATTAGCCGGGCGTGATGGTGGGCGCCTGTAGTCCCAGCTACACGGGAGGCTGAGACAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCACCACTGCACTGCAGCCTGGGCGACAGAGCGAGACTCTGTCTCAAAAAAAAAAAAGAAAGAAAAAAAACTAGCTAGCTAG, where SEQ ID NO: 44 is BRCA1-1-Alu4 without non-homologous sequences. The sequence was obtained by chemical synthesis and designated BRCA1-2-Alu4. SEQ ID NO: 45 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAGACTAAAAATACAAAAAATTAGCCGGGCGTGATGGTGGGCGCCTGTAGTCCCAGCTACACGGGAGGCTGAGACAGGAGAATGGCGTGAACCCGGGAGGCGGAGCTTGCAGTGAGCCGAGATCGCACCACTGCACTGCAGCCTGGGCGACAGAGCGAGACTCTGTCTCAAAAAAAAAAAAGAAAGAAAAAAAACTAGCTAGCTAG, where SEQ ID NO: 45 is BRCA1-1-Alu5 with no non-homologous sequence. The sequence was obtained by chemical synthesis and designated BRCA1-2-Alu5. BRCA1-1-Alu3, BRCA1-1-Alu4, BRCA1-1-Alu5, BRCA1-2-Alu3, BRCA1-2-Alu4, and BRCA1-2-Alu5 were each inserted into the plasmid vector pBS-L1PA1-CH-mneo to construct the plasmids pBS-L1PA1-CH-mneo-BRCA1-1-Alu3, pBS-L1PA1-CH-mneo-BRCA1-1-Alu4, pBS-L1PA1-CH-mneo-BRCA1-1-Alu5, pBS-L1PA1-CH-mneo-BRCA1-2-Alu3, pBS-L1PA1-CH-mneo-BRCA1-2-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-2-Alu5, as follows. BRCA1-1-Alu3, BRCA1-1-Alu4, BRCA1-1-Alu5, BRCA1-2-Alu3, BRCA1-2-Alu4, BRCA1-2-Alu5, and the plasmid pBS-L1PA1-CH-mneo were digested separately and the reaction systems are shown in Table 39: Table 39 Digestion reaction system The reaction was incubated at 37°C for 1 hour, then at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested BRCA1-1-Alu3, BRCA1-1-Alu4, BRCA1-1-Alu5, BRCA1-2-Alu3, BRCA1-2-Alu4, and BRCA1-2-Alu5 were ligated with the plasmid vector pBS-L1PA1-CH-mneo, and the reaction system is shown in Table 40: Table 40 Linkage reaction systems The reaction conditions were incubation at 16°C for 16 hours, followed by incubation at 70°C for 10 minutes to inactivate the ligase. The resulting plasmids were electrophoresed and purified to obtain pBS-L1PA1-CH-mneo-BRCA1-1-Alu3, pBS-L1PA1-CH-mneo-BRCA1-1-Alu4, pBS-L1PA1-CH-mneo-BRCA1-1-Alu5, pBS-L1PA1-CH-mneo-BRCA1-2-Alu3, pBS-L1PA1-CH-mneo-BRCA1-2-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-2-Alu5. The plasmids were sequenced to confirm their integrity. The group transfected with pBS-L1PA1-CH-mneo-BRCA1-1-Alu3, pBS-L1PA1-CH-mneo-BRCA1-1-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-1-Alu5 was designated experimental group 15, while the group transfected with pBS-L1PA1-CH-mneo-BRCA1-2-Alu3, pBS-L1PA1-CH-mneo-BRCA1-2-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-2-Alu5 was designated control group 8. Each group contained three parallel plates, each consisting of a 6-well plate cultured with HeLa cells. For the transfection procedure, HeLa cells were passaged and spread onto 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each 6-well plate, 96 μg (32 μg of each plasmid) of the constructed plasmid was diluted with 300 μL of serum-free DMEM and mixed thoroughly. At the same time, 120 μL of Entranster-H4000 reagent was diluted with 300 μL of serum-free DMEM, mixed thoroughly, and left at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and left at room temperature for 15 minutes to form the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM medium containing 10% fetal bovine serum per well for transfection. When the cells reached approximately 90% confluence, the above procedure was repeated after passage. Once the cells reached approximately 90% confluence, they were removed for subsequent procedures. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 9: the upstream primer sequence is shown in SEQ ID NO: 46: 5'-CCCCTTTATCTCCTTCTG-3'; the downstream primer sequence is shown in SEQ ID NO: 47: 5'-ATTTCTCCCATTCCACTT-3'. The upstream primer sequence of primer pair 9 is present only on the genome, not on the plasmid, and is located downstream of the 3'-terminal sequence of the sequence to be deleted in the intact BRCA1 gene, and the downstream primer sequence of primer pair 9 is located only on the genome, not in the plasmid, downstream of the 3'-terminal sequence of the sequence to be deleted in the intact BRCA1 gene. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 41. Table 41 qPCR reaction system JPEG2024504592000048.jpg81107 Cell DNA templates were extracted from the control group 8 and experimental group 15 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 9: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 46°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. After observing the exponential growth cycles in the amplification curves of GAPDH and the detection of the insertion of the downstream sequence of the 3'-terminal sequence of the sequence to be deleted and confirming that they are nearly parallel, the obtained data are analyzed. -ΔΔCt The results are shown in Table 42. The PCR products were confirmed to be correct by sequencing. Table 42 Results of primer pair 9 (n=3, x±s) JPEG2024504592000049.jpg132169The relative copy number (N / A calculated at 40.00) of experimental group 15 was significantly lower than that of control group 8, and the experimental results were statistically significant (P<0.05). This means that the gene portion at the CNV end of experimental group 15 has a lower copy number in the downstream sequence of the corresponding intact gene, indicating that the insertion of non-homologous sequences at the CNV end prevents the extension of the gene portion downstream of the CNV end. Conclusion: We show that the insertion of non-homologous sequences into the ends of CNVs prevents the corresponding CNV extension. Example 12: CNV terminal truncation (shortening) The 3' sequence of one of the introns of the BRCA1 gene (located in sequence number 39 in Example 11) was selected, and sequences were synthesized in the following order: 3' terminal sequence of the sequence to be deleted + non-homologous sequence + upstream sequence immediately adjacent to the 5' end of the sequence to be deleted + partial Alu sequence. NheI digestion sites and protecting bases were added to both ends, and the sequences are shown in sequence numbers 48, 49, and 50. The sequence of SEQ ID NO: 48 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG GGGGGG The shadow sequence is a partial Alu3 sequence, with NheI digestion sites and protective bases (bold italics) at both ends. Between the shadow sequence and the non-homologous sequence is the upstream sequence immediately adjacent to the 5' end of the sequence to be deleted. The shadow sequence is a partial Alu3 sequence, Alu3. The sequence was obtained by chemical synthesis and designated BRCA1-3-Alu3. SEQ ID NO: 49 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG GGGGGG The shadow sequence is a partial Alu4 sequence. ... The sequence was obtained by chemical synthesis and designated BRCA1-3-Alu4. SEQ ID NO:50 is: CTAGCTAGCTAGACTAACATCATTTGGAAATAATTTCATGGGCATTAATTGCATGAATGTGGTTAGATTAAAAGGTGTTCAGCTAGAACTTGTAGTTCCATACTAGGTGATTTCAATTCCTGTGCTAAAATTAATTTGTATGATATATTTTCATTTAATGGAAAGCTTCTCAAAGTATTTCATTTTCTTGGTGCCATTTATCGTTTTTGAAG GGGGGGThe shadow sequence is a partial Alu5 sequence, with NheI digestion sites and protective bases (italicized bold) at both ends. The shadow sequence is immediately upstream of the 5' end of the sequence to be deleted. The shadow sequence is a partial Alu5 sequence. The sequence was obtained by chemical synthesis and designated BRCA1-3-Alu5. BRCA1-3-Alu3, BRCA1-3-Alu4, and BRCA1-3-Alu5 were each inserted into the plasmid vector pBS-L1PA1-CH-mneo to construct the plasmids pBS-L1PA1-CH-mneo-BRCA1-3-Alu3, pBS-L1PA1-CH-mneo-BRCA1-3-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-3-Alu5 as follows. BRCA1-3-Alu3, BRCA1-3-Alu4, BRCA1-3-Alu5, and the plasmid pBS-L1PA1-CH-mneo were digested separately, and the reaction sequences are shown in Table 43: Table 43 Digestion reaction system The reaction was incubated at 37°C for 1 hour, then at 65°C for 20 minutes to inactivate the endonuclease enzyme. The digested product was then electrophoresed and collected. The digested BRCA1-3-Alu3, BRCA1-3-Alu4 and BRCA1-3-Alu5 were ligated with the plasmid vector pBS-L1PA1-CH-mneo, and the reaction system is shown in Table 44: Table 44 Linkage reaction systems The reaction conditions were incubation at 16°C for 16 hours, followed by incubation at 70°C for 10 minutes to inactivate the ligase. The resulting plasmids were electrophoresed and purified to obtain pBS-L1PA1-CH-mneo-BRCA1-3-Alu3, pBS-L1PA1-CH-mneo-BRCA1-3-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-3-Alu5. The plasmids were sequenced to confirm their integrity. The group transfected with pBS-L1PA1-CH-mneo-BRCA1-3-Alu3, pBS-L1PA1-CH-mneo-BRCA1-3-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-3-Alu5 was designated experimental group 16, while the group transfected with pBS-L1PA1-CH-mneo-BRCA1-1-Alu3, pBS-L1PA1-CH-mneo-BRCA1-1-Alu4, and pBS-L1PA1-CH-mneo-BRCA1-1-Alu5 was designated control group 9. Each group contained three parallel plates, each consisting of a 6-well plate cultured with HeLa cells. For the transfection procedure, HeLa cells were passaged and spread onto 6-well plates. The following day, Entranster-H4000 transfection reagent was used for transfection. For each 6-well plate, 96 μg (32 μg of each plasmid) of the constructed plasmid was diluted with 300 μL of serum-free DMEM and mixed thoroughly. At the same time, 120 μL of Entranster-H4000 reagent was diluted with 300 μL of serum-free DMEM, mixed thoroughly, and left at room temperature for 5 minutes. The two liquids were then combined, mixed thoroughly, and left at room temperature for 15 minutes to form the transfection complex. The transfection complex was then added to a 6-well plate containing 2 ml of DMEM medium containing 10% fetal bovine serum per well for transfection. When the cells reached approximately 90% confluence, the above procedure was repeated after passage. Once the cells reached approximately 90% confluence, they were removed for subsequent procedures. Extraction of transfected cell DNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Then, extract the cell DNA according to the instructions of the blood / cell / tissue genomic DNA extraction kit, and measure the DNA concentration using a UV spectrophotometer. qPCR testing: The GAPDH gene is used as an internal reference gene because it does not contain Alu sequences and its copy number is stable. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 10: the upstream primer sequence is shown in SEQ ID NO: 51: 5'-GCTTTCTCAGGGCTCTTT-3'; the downstream primer sequence is shown in SEQ ID NO: 52: 5'-GCACCATCTCGGCTCACT-3'. The upstream primer sequence of primer pair 10 is located on the sequence to be deleted and is not present in the plasmid but only in the genome, while the downstream primer sequence of primer pair 10 is located on the sequence to be deleted and is not present in the plasmid but only in the genome. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 45. Table 45 qPCR reaction system JPEG2024504592000052.jpg80107 Cell DNA templates were extracted from the control group 9 and experimental group 16 after transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 10: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 49°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. After observing the exponential growth cycles of the amplification curves of GAPDH and the sequence to be deleted and confirming that they are nearly parallel, the data obtained are analyzed by 2 -ΔΔCt The results are shown in Table 46. The PCR products were confirmed to be correct by sequencing. Table 46 Results of primer pair 10 (n=3, x±s) JPEG2024504592000053.jpg74169JPEG2024504592000054.jpg60169The relative copy number (N / A calculated at 40.00) in experimental group 16 was significantly lower than that in control group 9, and the experimental results were statistically significant (P<0.05), indicating that in experimental group 16, the deleted sequence was deleted and the gene subsequence at the end of the CNV was reduced. Example 11 shows that a non-homologous sequence can be inserted into the CNV end to prevent the CNV end from continuing to extend. However, Example 12 shows that the sequence of the gene portion at the CNV end in the experimental group is significantly smaller than that in the control group, indicating that the CNV end is clipped while the CNV is shortened, demonstrating that the CNV end can be modified by the related method of the present invention. Therefore, in the gene editing method of the present invention, multiple or all CNVs in the genome can also be modified by changing the guide sequence upstream of the insertion point (i.e., the target site upstream sequence) (when gene editing is performed at the CNV end, the target site upstream sequence and the gene portion sequence at the CNV end are identical). Example 13 Corresponding gene expression changes after CNV terminal clipping Total RNA is extracted from cells of experimental group 16 and control group 9 in Example 12: Extraction of transfected cell RNA: After aspirating the cell culture medium, rinse the cells twice with PBS. Add an appropriate amount of 0.25% trypsin for digestion and digest for a total of 20 minutes at 37°C, pipetting (aspirating and blowing) 15 times every 5 minutes. When the cells are in suspension, add complete medium containing serum to terminate the reaction. Transfer the cell-containing solution to an RNase-free centrifuge tube and centrifuge at 300g for 5 minutes. Collect the pellet and aspirate all the supernatant. Perform total RNA extraction according to the instructions of the magnetic bead method tissue / cell / blood total RNA extraction kit. Extraction of mRNA from total RNA: The concentration of the previously extracted total RNA was detected by UV spectrophotometer, 1000 ng of total RNA was diluted to 50 µL with nuclease-free ddHO, mRNA was extracted according to the instructions of the TIANSeq mRNA Capture Kit, and an aliquot of the sample was taken to detect the mRNA content (the above experimental procedure was repeated several times to obtain enough mRNA for transfection). Reverse transcription synthesis of cDNA template: The cDNA is synthesized according to the instructions of the FastKing cDNA First Strand Synthesis Kit, and the concentration of the synthesized cDNA is determined by UV spectrophotometer for subsequent testing. qPCR testing: The GAPDH gene is used as an internal reference gene because its expression is relatively stable in various tissues. The upstream primer sequence for the GAPDH gene is shown in SEQ ID NO:7, and the downstream primer sequence is shown in SEQ ID NO:8. Primer pair 11: The upstream primer sequence is shown in SEQ ID NO: 53: 5'-CAGAGGACAATGGCTTCCATG-3'; the downstream primer sequence is shown in SEQ ID NO: 54: 5'-CTACACTGTCCAACACCCACTCTC-3'. The upstream primer sequence of primer pair 11 is located on the BRCA1 gene and is not present in the plasmid but only in the genome, and the downstream primer sequence of primer pair 11 is located on the BRCA1 gene and is not present in the plasmid but only in the genome. The above primers are obtained by chemical synthesis. The qPCR reaction system is shown in Table 47. Table 47 qPCR reaction system JPEG2024504592000055.jpg80107 Cell DNA template was used to synthesize cDNA from mRNA extracted from 9 control and 16 experimental groups after co-transfection. Reactions were prepared on ice, capped, mixed gently, and briefly centrifuged to ensure all components were at the bottom of the tube. Each 6-well plate cell sample was run in triplicate. qPCR reaction cycle: Primer pair 11: Pre-denaturation at 95°C for 15 minutes; 40 cycles of denaturation at 95°C for 10 seconds, annealing at 55°C for 20 seconds, and extension at 72°C for 20 seconds. GAPDH primers were reacted under the same conditions. After observing the exponential growth cycles in the amplification curves of GAPDH and BRCA1 expression and confirming that they were nearly parallel, the data were analyzed. -ΔΔCtThe results are shown in Table 48. The PCR products were confirmed to be correct by sequencing. Table 48 Results of primer pair 11 (n=3, x±s) JPEG2024504592000056.jpg82169JPEG2024504592000057.jpg60169The relative expression of the BRCA1 gene in experimental group 16 was lower than that in control group 9, which was statistically significant (P<0.05), indicating that the CNV terminal clipping of the BRCA1 gene led to a decrease in its expression. Conclusion: We found that the expression of the BRCA1 gene was decreased, indicating that CNV editing can affect the transcription of the corresponding gene, potentially affecting protein expression and the state of cells, tissues, and organisms. As can be seen from Examples 11-13, the present invention allows for the fixation, extension, and clipping of CNV ends, while simultaneously affecting cellular gene transcription and protein expression. Furthermore, CNVs change depending on physiological processes, such as embryonic and ontogenetic development and tumorigenesis, and vary between cells, tissues, and individuals. Therefore, editing CNVs can also alter the state of the corresponding cells, tissues, and organisms. As can be seen from the above embodiments, the present invention uses retrotransposons, which are widely present in eukaryotes, and their reverse transcription function to edit genomes. The involved SINE and LINE sequences and associated proteins are widely present in normal organisms. Target fragments can be integrated into the genome, and corresponding fragments can be deleted and replaced without generating double-strand breaks, resulting in more precise target sequence recognition and cleavage. Because there are no double-strand breaks, there is no need to worry about the risk of genomic double-strand DNA breaks or the introduction of unexpected random sequences. Taking the Alu element sequence in SINE and its function corresponding to LINE-1 in LINE as an example, Alu elements and LINE-1 are widely distributed in the genome of primates, and in a specific embodiment, the inserted sequence is guided to an insertion site (target site) on the genome depending on the sequences on both sides of the inserted sequence (the upstream sequence of the target site and the downstream sequence of the target site), Furthermore, ORF2p can slide smoothly from the 3' end of the carrier nucleic acid to the shearing site for single-strand cleavage on the genome only under conditions where the upstream sequence of the target site is perfectly matched, thereby greatly improving targeting accuracy and avoiding unexpected cleavage. This targeting accuracy is theoretically higher than that of other currently available gene editing technologies. Furthermore, by generating the necessary RNAs and corresponding proteins, such as ORF1p and ORF2p, in vitro, target sequences, genes, and genomes can be modified via the RNA or RNP pathway without introducing DNA fragments or entering the nucleus during transfection. The nuclear localization function of ORF1p (and ORF2p) allows transfected RNAs and proteins to enter the nucleus, which is beneficial for editing cells that are difficult to manipulate due to the difficulty of vector entry into the nucleus. At the same time, this invention can also edit genomic CNVs, which can be increased, decreased, or stabilized (i.e., cannot remain unchanged) through this method. Since CNVs can directly affect protein expression, manipulation of CNVs can change or stabilize the expression and state of corresponding cells, tissues, organs, or organisms. Because various types of SINEs and LINEs, such as various MIRs and LINE-2s, which are homologous and functionally similar to Alu elements and LINE-1s, are widely distributed in eukaryotes, this invention can also be applied to other eukaryotic systems. Unlike other gene editing technologies, the relevant mechanisms used in the present invention exist in normal organisms, and do not introduce foreign mechanisms and systems, thereby reducing the impact on the recipient system for gene editing.Since the present invention does not introduce foreign systems such as prokaryotic proteins and does not cause double-strand breaks, it is easier to apply the present invention to clinical settings than other existing gene editing technologies.

Claims

1. Use of a gene transcription framework for inserting, deleting, or substituting DNA sequences in any region of a genome by transcription, comprising: The gene transcription framework comprises, in the 5' to 3' direction, an upstream sequence of a target site, a sequence to be inserted, and a downstream sequence of the target site; the gene transcription framework is a DNA sequence that can be transcribed by RNA polymerase I, RNA polymerase II or RNA polymerase III; a target site upstream sequence or its complementary sequence in a transcription product of the gene transcription framework or a transformation product of the transcription product is capable of hybridizing with a corresponding target site upstream sequence or its complementary sequence in a cell genome, a target site downstream sequence or its complementary sequence is capable of hybridizing with a corresponding target site downstream sequence or its complementary sequence in a cell genome, the target site upstream sequence and the target site downstream sequence are directly connected in a corresponding target gene sequence in the genome, and a site between the target site upstream sequence and the target site downstream sequence in the target gene sequence on the genome is a target site for a sequence to be inserted; The use of a gene transcription framework, characterized in that the cell is a eukaryotic cell.

2. A vector system, comprising one or more types of vectors, 1) one or more gene transcription frameworks according to claim 1; 2) one or more short interspersed element sequences and / or partial short interspersed element sequences and / or similar short interspersed element sequences; and / or one or more of 3) long interspersed element sequences and / or ORF1p coding sequences and / or ORF2p coding sequences; Including, wherein the components 1), 2) and / or 3) are located in the same or different vectors of the vector system; the vector comprises one or more promoters, the promoters being an RNA polymerase I promoter, an RNA polymerase II promoter or an RNA polymerase III promoter, and located upstream of the components 1), 2) and / or 3); A vector system, characterized in that the vector system is mediated by a DNA, RNA and / or RNP pathway.

3. 3. The vector system according to claim 2, wherein component 1), if present, is located in the same or different vectors of the vector system; component 2), if present, is located in the same or different vectors of the vector system; and component 3), if present, is located in the same or different vectors of the vector system.

4. The vector system according to claim 2, characterized in that the vector is a eukaryotic expression vector, a prokaryotic expression vector, a viral vector, a plasmid vector, an artificial chromosome, a phage vector, or a cosmid vector.

5. The vector system according to claim 2, characterized in that the vector is an expression vector, a cloning vector, a sequencing vector, a transformation vector, a shuttle vector or a multifunctional vector.

6. A vector system as described in claim 2, characterized in that when 1) and 2) are located in the same vector, the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence are located downstream of the gene transcription framework and are directly or indirectly connected to the gene transcription framework and the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence, and in the case of a direct connection, the gene transcription framework shares one promoter with the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence, and in the case of an indirect connection, the gene transcription framework shares one promoter or does not share one promoter with the short interspersed element sequence, and / or partial short interspersed element sequence, and / or similar short interspersed element sequence.

7. A vector system as described in claim 2, characterized in that when 1) and 3) are located in the same vector, the one or more long interspersed element sequences, and / or the one or more ORF1p coding sequences, and / or the one or more ORF2p coding sequences are located upstream and / or downstream of the gene transcription framework, and the gene transcription framework is directly or indirectly connected to the one or more long interspersed element sequences, and / or the one or more ORF1p coding sequences, and / or the one or more ORF2p coding sequences, and in the case of direct connection, the gene transcription framework shares a promoter with the one or more long interspersed element sequences, and / or the one or more ORF1p coding sequences, and / or the one or more ORF2p coding sequences, and in the case of indirect connection, the gene transcription framework shares a promoter or does not share a promoter with the one or more long interspersed element sequences, and / or the one or more ORF1p coding sequences, and / or the one or more ORF2p coding sequences.

8. A vector system according to claim 2, wherein, when 1), 2) and 3) are located on the same vector, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are located downstream of the gene transcription framework and / or downstream of the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, and when the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are located downstream of the gene transcription framework, the long interspersed element sequence and and / or the ORF1p coding sequence and / or the ORF2p coding sequence are located upstream of said gene transcription framework, and / or said long interspersed element sequence and / or the ORF1p coding sequence and / or the ORF2p coding sequence are located downstream of said short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence, and said short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are located downstream of said long interspersed element sequence and / or the ORF1p coding sequence and / or the ORF2p coding sequence. When the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence are located downstream of the gene transcription framework, and when the gene transcription framework, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are directly or indirectly connected to the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, the gene transcription framework, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are directly or indirectly connected to the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, or partial short interspersed element sequences and / or similar short interspersed element sequences share a promoter with said long interspersed element sequences and / or ORF1p coding sequence and / or ORF2p coding sequence and are indirectly connected thereto, said gene transcription framework, said short interspersed element sequences and / or partial short interspersed element sequences and / or similar short interspersed element sequences share a promoter or do not share a promoter with said long interspersed element sequences and / or ORF1p coding sequence and / or ORF2p coding sequence,Vector system.

9. A vector system according to claim 2, wherein when 2) and 3) are located on the same vector, the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence are located upstream and / or downstream of the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence, and the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence are directly or indirectly connected to the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, and directly or indirectly connected to the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence. A vector system characterized in that, when connected, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence share a promoter with the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence, and when indirectly connected, the short interspersed element sequence and / or partial short interspersed element sequence and / or similar short interspersed element sequence share a promoter or do not share a promoter with the long interspersed element sequence and / or ORF1p coding sequence and / or ORF2p coding sequence.

10. A method for editing a genome sequence, comprising: 1) selecting a site into which a target gene to be edited is to be inserted from the genome, and determining the upstream and downstream sequences of the site into which the target gene is to be inserted on both sides of the insertion site; 2) producing a vector system according to any one of claims 2 to 9; 3) transforming or transfecting a cell, tissue, or organism with the vector system to perform gene / genome editing; A method for editing a genome sequence, comprising:

11. Use of a vector system mediated by a DNA, RNA and / or RNP pathway as described in any one of claims 2 to 9 in the insertion, deletion or replacement of a DNA sequence in any region of a genome.

12. The use according to claim 11, characterized in that the DNA sequence is one or more CNV sequences, CNV end sequences, short interspersed element sequences, partial short interspersed element sequences, similar short interspersed element sequences, long interspersed element sequences, partial long interspersed element sequences, ORF1p coding sequences and / or ORF2p coding sequences.

13. Use of a vector system described in any one of claims 2 to 9 as a medicine for the prevention and / or treatment of cancer, gene-related genetic diseases, and neurodegenerative diseases.

14. The use according to claim 13, wherein the cancer is selected from the group consisting of glioma, breast cancer, cervical cancer, lung cancer, gastric cancer, colon cancer, duodenal cancer, leukemia, prostate cancer, uterine cancer, thyroid cancer, lymphoma, pancreatic cancer, liver cancer, melanoma, skin cancer, pituitary tumor, germ cell tumor, meningioma, meningeal cancer, glioblastoma, various astrocytomas, various oligodendrogliomas, astrocytomas, various ependymomas, choroid plexus papilloma, choroid plexus carcinoma, chordoma, neuroblastoma, various ganglioneuromas, olfactory neuroblastoma, sympathetic neuroblastoma, pineal cell tumor, pineoblastoma, medulloblastoma, trigeminal neurinoma, acoustic neuroma, jugular vein tumor, jugular globus tumor, hemangioplexioma, craniopharyngioma, granular cell tumor and / or granulosa cell tumor.

15. The use of claim 13, wherein the gene-related genetic disease is Huntington's disease, fragile X syndrome, phenylketonuria, pseudohypertrophic progressive muscular dystrophy, mitochondrial encephalomyopathy, spinal muscular atrophy, Parkinsonian superposition syndrome, Parkinsonism, Parkinson's disease, albinism, red-green color blindness, red-green color dysphoria, achondroplasia, alkaptonuria, congenital deafness, thalassemia, sickle cell anemia, hemophilia, epilepsy associated with genetic changes, myoclonus, dystonia, stroke and schizophrenia, vitamin D resistant rickets, familial colonic polyposis, hereditary nephritis.

16. The use according to claim 13, wherein the neurodegenerative disease is Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar ataxia, multiple system atrophy, primary lateral sclerosis, Pick's disease, frontotemporal dementia, dementia with Lewy bodies, or progressive supranuclear palsy.