264 results about "Recombinase" patented technology

Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity of RNA-programmable endonucleases, such as Cas9, or for controlling the activity of proteins comprising a Cas9 variant fused to a functional effector domain, such as a nuclease, nickase, recombinase, deaminase, transcriptional activator, transcriptional repressor, or epigenetic modifying domain. For example, the inventive proteins provided comprise a ligand-dependent intein, the presence of which inhibits one or more activities of the protein (e.g., gRNA binding, enzymatic activity, target DNA binding). The binding of a ligand to the intein results in self-excision of the intein, restoring the activity of the protein.

The invention belongs to the technical field of biology, and particularly relates to an SLC44A1 gene and / or SLC44A5 gene knockout animal model as well as a construction method and application thereof. The construction method of the animal model comprises the following steps: deleting exon regions from the fourth exon region to the thirteenth exon region of the SLC44A1 gene and / or the second exon region of the SLC44A5 gene in mouse brain oligodendroglia cells in a high-efficiency manner by utilizing gene editing and a Cre-loxP recombinase system, so as to realize the knockout of a target gene. The animal model constructed by the invention can be used for researching the regulation mechanism of the choline transporters (SLC44A1 and SLC44A5) on myelin sheath formation, deeply knowing the biological functions of the choline transporters (SLC44A1 and SLC44A5), simulating the occurrence and development processes of human related diseases, revealing the pathogenesis of the related diseases, providing a new thought for prevention and treatment of brain myelin sheath dysplasia, and providing a new foundation for the prevention and treatment of brain myelin sheath dysplasia. And an effective drug or a treatment mode aiming at the disease is screened by utilizing a gene knockout animal model, so that a new choice is provided for clinical treatment of the disease.

The invention discloses a CRISPR / Cas12a-RPA (clustered regularly interspaced short palindromic repeats / CRISPR associated protein 12a-recombinase polymerase amplification)-based sequence combination for rapidly detecting mouse hepatitis virus and a kit thereof, and belongs to the technical field of biological detection The sequence combination comprises an RPA primer pair sequence, a crRNA sequence and an ssDNA probe sequence; the kit comprises an RPA primer pair sequence, a crRNA sequence and an ssDNA probe sequence, the kit further comprises a mouse hepatitis virus negative control standard substance, RNase-free water, an RPA reaction buffer solution, an NEB buffer solution, Lba Cas12a nuclease, dry powder RPA reaction microspheres containing recombinase polymerase and magnesium acetate with the concentration of 280 mM. The kit can be used for rapidly detecting the mouse hepatitis virus. The primer sequence combination provided by the invention is good in conservative property and specificity and relatively high in sensitivity, and rapid detection of the mouse hepatitis virus can be realized.

The invention discloses a method for culturing CIK (Cytokine-Induced Killer) cells and application thereof. The method comprises the following steps: recovering cord blood, mixing and centrifuging the recovered cord blood and a buffer solution, re-suspending and precipitating, centrifuging again to obtain cord blood mononuclear cells, inoculating the cord blood mononuclear cells into an activation culture medium, and culturing to obtain the CIK cells, the buffer solution contains L-proline, human recombinant DNA enzyme and angelica polysaccharide. According to the method, the mononuclear cells can be separated from the umbilical cord blood preserved by liquid nitrogen, the utilization rate of the umbilical cord blood which can only be discarded due to insufficient cells is increased, the traditional lymphocyte separation process is optimized, the recovery rate of the mononuclear cells is increased, and the cost is reduced. The method provided by the invention can be used for culturing dozens to ten billions of CIK cells from a small amount of cryopreserved cord blood, and has huge advantages in the field of CIK cell culture and application.

The invention discloses a construction method and application of a mouse model for specifically tracing heart valve cells, and belongs to the technical field of animal model construction. The model construction method comprises the following steps: (1) constructing a Wid1-Cre hybrid mouse before inserting a Cre recombinase coding gene into a termination codon of a mouse Wid1 gene; and (2) hybridizing the Wid1-Cre hybrid mouse with the Rosa26-tdTomato homozygous report mouse, so as to obtain the double transgenic mouse which simultaneously carries the Wid1-Cre gene and the Rosa26-tdTomato gene. According to the invention, on the basis of the expression characteristic that Wif1 is only limited to a valve area in the heart, Wif1-Cre is constructed; according to the Rosa26-tdTomato mouse model, specific tracing of heart valve cells is achieved, and an important experimental tool is provided for mechanism research of heart valve diseases and development of therapeutic drugs.

The invention relates to the technical field of biology, and particularly discloses a method for detecting corn leukoderma pathogenic bacteria based on RPA-LFD (Recombinase Polymerase Amplification-Lateral Flow Dipstick), which comprises the following steps: a recombinase polymerase amplification RPA reaction system comprising an upstream primer PA-F2, a biotin-labeled downstream primer PA-R3-biotin and an nfo probe PA3-nfo-2; the lateral flow chromatography analysis LFD test strip is used for detecting an amplification product; a reaction buffer solution and a dilution buffer solution; a rapid and visual corn leaf spot detection method is developed by using a recombinase-mediated isothermal amplification (PRA) technology and combining an nfo probe with a lateral flow chromatography analysis and diagnosis method, and finally, an efficient and rapid corn leaf spot detection, prevention and control system is established by popularizing and applying a novel visual detection method in a field.

The invention belongs to the technical field of biological medicine, and particularly relates to an engineered cell microvesicle and a delivery system based on the engineered cell microvesicle, the system realizes efficient preparation of 1-5 [mu] m cell microvesicles, and the cell microvesicles have a large space volume and can be used for preparing the cell microvesicles. The carrier can be used for loading and delivery of target protein, polypeptide and recombinase which are specifically expressed in mother cells. The system transfects mother cells through lentivirus transfection or plasmid transfection to further produce cell microvesicles, and the microvesicles can load more goods and inherit membrane proteins of the mother cells, and can also effectively load intracellular proteins to realize effective delivery. By virtue of good structural stability, high immunogenicity and excellent biocompatibility, the cell microvesicle reduces systematic toxic and side effects of a traditional carrier, is expected to become an effective drug delivery system, and has great application potential in the field of gene therapy.

SpRYc is a grafted ScCas9++-SpRY chimeric Cas9 possessing minimal 5′-NNN-3′ PAM specificity. SpRYc comprises the N-terminus (residues 1-1119) of ScCas9++ (Sc++), including the flexible loop, followed by the region of SpRY (residues 1111-1368) spanning its PAM-interacting domain mutations. Methods of altering gene expression include use of SpRYc in complex with guide RNA in a CRISPR-Cas9 system.

The invention discloses a preparation method of recombinant catechol-2, 3-dioxygenase and an application of the recombinant catechol-2, 3-dioxygenase. According to the invention, a novel catechol-2, 3-dioxygenase group C23O927 is identified from a constructed metagenome library, and the catechol-2, 3-dioxygenase group C23O927 is a key enzyme for catalyzing degradation of polycyclic aromatic hydrocarbon pollutants. A C23O927-pET32a (+) recombinant expression vector is constructed and converted into host bacteria, heterologous expression of target protein is achieved, and the prepared recombinase belongs to acid enzymes and has good resistance to high temperature, acid, alkali, high-concentration metal ions, organic solvents and surfactants. The recombinase provided by the invention can be used in an acidic environment, has very good enzymatic activity, can catalytically degrade various catechol compounds, can be better used for treating polycyclic aromatic hydrocarbon polluted environments, and has important significance for developing more products which can tolerate various extreme environments and efficiently catalyze degradation of polycyclic aromatic hydrocarbons.

The invention relates to the technical field of technical gene detection, in particular to a recombinase and recognition site screening method and system based on structure classification prediction.The method comprises the steps that to-be-matched recombinase is obtained for structure resolution prediction.The functional domain of the to-be-matched recombinase is obtained; searching in a first database according to the functional domain, searching candidate recombinase of which the similarity meets a preset condition, and screening to obtain optimal recombinase; determining an interaction region of the current large serine recombinase and the target DNA sequence from a second database, and sequentially obtaining interaction amino acid sequences corresponding to the preferable recombinase; and matching with the amino acid characteristic sequence of the current large serine recombinase in the second database to obtain the target large serine recombinase with the highest similarity in the preferred recombinase and the corresponding recognition site sequence. According to the method, the structure of the large protein is subjected to structure comparison mining and splitting, and recognition site prediction is performed on interaction region amino acid characteristics, so that the research and development period is shortened, and the research and development cost is saved.

The invention relates to the field of molecular diagnosis and nucleic acid amplification, in particular to a recombinase polymerization amplification method based on Twinkle helicase enhancement and application of the recombinase polymerization amplification method. According to the invention, Twinkle helicase is introduced into an RPA system for the first time, and primer combination and template unlinking are assisted by utilizing ATP-dependent unlinking activity and DNA annealing activity of the Twinkle helicase, so that the amplification efficiency and the detection sensitivity are remarkably improved. The method is suitable for plasmid DNA, a high GC template and a clinical nucleic acid sample, and can be used for constructing an RPA kit containing Twinkle, so that the application of the RPA kit in pathogen detection, molecular diagnosis and on-site rapid detection is expanded. According to the invention, the defects of the existing RPA technology in sensitivity and stability are overcome, and a new solution is provided for developing a novel nucleic acid self-detection and field detection tool.

The present disclosure relates to enzymes, compositions and methods for performing conditional homologous recombination of a targeted DNA molecule or genome by using modified proteins comprising nucleic acid DNA binding proteins with protease-conditional recombinase (hereinafter “ProRec”) activity. Namely a new cre recombinase pro enzyme, compositions / kit and sensors comprising the cre recombinase pro enzyme of the present disclosure. Namely, a cre recombinase pro-enzyme, for identifying and quantifying proteolytic activity in a disease or an infection, comprising intein mediated circularization, a flip-excision cassette comprising an output reporter and / or effector protein; a linker sequence cleavable by a protease.

The invention belongs to the technical field of biological engineering, and provides curcumin / dihydrocurcumin reductase MaCurA, a coding gene and application. The reductase can efficiently catalyze 1, 7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-diketone to synthesize tetrahydrocurcumin, an efficient solution is provided for industrial preparation of tetrahydrocurcumin, and experiments prove that the recombinase can catalyze the synthesis of tetrahydrocurcumin in the catalysis of 1, 7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-diketone. In a reaction system for preparing tetrahydrocurcumin from 2, 5-diketone, actually measured enzyme activity is stabilized at 3.38 U / mg level, and compared with a traditional chemical synthesis method, the production efficiency is remarkably improved.

The invention discloses a method for detecting high sensitivity of monkey pox virus by a one-pot method RPA-CRISPR based on temperature control, and the method comprises the following steps: establishing a temperature-controlled RPA and CRISPR-Cas12b combined detection method, which is called as CRATS for short, by utilizing the reaction temperature difference between RPA amplification and CRISPR-Cas12b cutting; the reaction process in a tube system is regulated and controlled by stages through a temperature switching strategy: target amplification is completed at 37 DEG C firstly, then the temperature is raised to 60 DEG C, Cas12b cutting is started, so that sequential separation of amplification and cutting reactions is realized, and finally detection is realized through a fluorescence signal released by molecular beacon breakage. The problem that Cas enzyme cutting activity interferes with the amplification process in a traditional one-pot method reaction is solved, the detection sensitivity and specificity are improved, and a new technical means is provided for on-site rapid detection of the monkey pox virus.

The invention discloses Bxb1 recombinase with improved activity and application of the Bxb1 recombinase. The invention provides a recombinase, and the amino acid sequence of the recombinase comprises one or more amino acid substitutions relative to the amino acid sequence shown in SEQ ID NO: 1. The recombinase provided by the invention can act on large-fragment DNA in a genome, so that the activity of a recombinase genome editing system is improved, and the application scene and selection of recombinase genome editing are enriched.

The invention discloses a construction method for improving gene editing efficiency of a recombinant trichoderma reesei engineering strain, and belongs to the technical field of gene editing. In the research, the T.reesei CJ2095 is taken as an original strain, the gene ku80 is knocked out, and the gene editing efficiency is improved to 7.14% from 1.04% of that of a control strain. In order to realize the recycling of the screening marker of the orotidine-5 '-phosphate decarboxylase gene pyr4, the gene pyr4 is knocked out. In addition, homologous recombinase rad51 and rad52 genes of a T.reesei endogenesis are respectively overexpressed, and the overexpressed gene rad52, namely the homologous recombination efficiency of the engineering strain T.reesei CJ2095 delta ku80 delta pyr4: rad52, can be improved to 43.75%, which is 42 times of that of an original strain.

The invention discloses an artificially optimized cDNA (complementary deoxyribonucleic acid)-based high-expression trametes versicolor chitin endonuclease gene, a recombinant vector, recombinant bacteria, recombinant enzyme and a preparation method thereof. The optimized trametes versicolor chitin endonuclease gene, such as a sequence as shown in SEQ ID NO.1, is constructed into a pichia pastoris expression vector and then converted into a pichia pastoris expression strain; according to the method, recombinant bacteria expressing active trametes versicolor chitin endonuclease are obtained through high-Zeocin-resistance YPD plate screening, high-expression pichia pastoris transformants are further screened from high-Zeocin-resistance transformants, finally fermentation is carried out in a bioreactor, obtained supernate contains a large amount of recombinant active trametes versicolor chitin endonuclease, and the recombinant active trametes versicolor chitin endonuclease can be obtained under the high-density fermentation condition. The expression level of the recombinant trametes versicolor chitin endonuclease reaches 3.1 g / L, and the trametes versicolor chitin endonuclease is efficiently purified through ammonia sulfate fractional precipitation and ion exchange. The recombinase can hydrolyze colloidal chitin to generate chitin oligosaccharide, and has important application value and wide application prospect in the aspects of preparation of functional chitosan oligosaccharide, antibiosis and anti-tumor, agricultural and biological control, environmental protection, industrial catalysis and the like.

The application discloses a carbonyl reductase mutant and application thereof in preparation of (R)-8-chloro-6-hydroxyoctanoate compounds. The application relates to a mutant of carbonyl reductase with improved catalytic performance and stereoselectivity after rational molecular modification, a recombinant expression plasmid containing the mutant gene of the enzyme, a recombinant expression transformant, a preparation method of the recombinant enzyme, and application of the recombinant enzyme or the recombinant cell as a catalyst in asymmetric reduction of 8-chloro-6-carbonyloctanoate compounds (I) to prepare (R)-8-chloro-6-hydroxyoctanoate compounds (II), in particular, in an intermediate (R)-8-chloro-6-hydroxyoctanoate methyl ester of thioctic acid. The disclosed carbonyl reductase SsCR mutant has the advantages of high reaction efficiency and high catalytic activity, and exhibits the advantages of high substrate concentration, low catalyst consumption and short reaction time in the process of catalytic preparation of the compound (II).

The invention discloses polyethylene glycol terephthalate hydrolase PmPETase and application thereof.The hydrolase PmPETase is protein with the amino acid sequence shown in SEQ ID NO.1. An expression system suitable for the PmPETase is constructed, recombinase protein with catalytic activity is obtained, and the polyethylene glycol terephthalate hydrolase PmPETase can be used for preparing the polyethylene glycol terephthalate hydrolase PmPETase. The substrate specificity, the catalytic property and the potential plastic degradation capacity of the PET degrading enzyme are comprehensively analyzed, and a new theoretical basis and application reference are provided for development and utilization of the PET degrading enzyme and a high polymer material green treatment technology.

The invention relates to the technical field of water pollution and water body odor substance detection, in particular to a primer group, a kit and a method for detecting cyanobacteria generating 2-methylisoborneol in a water body based on a recombinase-mediated isothermal nucleic acid amplification (RAA) technology. The primer group is designed based on a mic gene conserved region of the 2-methylisoborneol-producing cyanobacteria, and comprises a forward primer, a reverse primer and a specific probe; the 5'end of the reverse primer is modified with a fluorescent marker, the 5 'end of the probe is modified with biotin, the 3' end of the probe is modified with a phosphorylation group, and nuclease cleavage sites are contained in the probe. The primer group disclosed by the invention can effectively amplify the mic gene of the 2-methylisoborneol, has relatively high specificity and sensitivity, has no cross reaction with cyanobacteria which does not generate the 2-methylisoborneol, and can be combined with lateral flow immunochromatography to be used for on-site rapid detection of the 2-methylisoborneol; the method is of great significance in early warning of excessive content of 2-methylisoborneol in the water body.

The present disclosure relates to production of cis-3-hexenol from unsaturated fatty acid, such as alpha linoleic acid and linoleic acid. The process of the present disclosure involves combinatorial and sequential action of recombinant enzymes, such as LOX, HPL and ADH for the production of cis-3-hexenol.

The present invention refers to a recombinant Ten Eleven Translocation 3 (TET3) enzyme, a nucleic acid molecule encoding the TET3 enzyme, a vector comprising the nucleic acid molecule and a host cell transformed or transfected with the vector, the use of the recombinant TET3 enzyme for sequencing of a nucleic acid substrate, a method for sequencing a nucleic acid substrate and a reagent for sequencing a nucleic acid substrate.

The invention relates to screening methods. In particular, it relates to a method of identifying a functional TCR from a library of particles, which displays a plurality of different T cell receptors (TCRs). The method may comprise a) exposing the library of particles to a target antigen to identify TCRs that bind to the target antigen, b) transfecting a plurality of T cells with nucleic acid encoding TCRs identified in step (a), c) exposing the plurality of transfected cells to the target antigen, and d) selecting cells having TCR activity in the presence of the target antigen. Transfection of the T cells is such that each T cell comprises a nucleic acid encoding a single TCR from the library of particles, the nucleic acid is integrated into the genome of the T cell at a single identical pre-defined locus, and the TCR is in a single chain format and comprises an alpha chain variable domain, a beta cain variable domain and a constant domain. The T cells do not express endogenous TCR. A T-cell comprising a recombinase mediated landing pad, wherein the T cell constitutively expresses TCR constant domain and does not express endogenous TCR, is also claimed.

The invention relates to a bacillus velezensis strain resistant to high temperature and alkali, and belongs to the technical field of microbial detoxification and feed safety. The bacillus velezensis B.26 strain with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.34475 is provided aiming at the problems that in the prior art, the thermal stability of zearalenone (ZEN) degrading enzyme is insufficient, the efficiency is low in an alkaline environment, and toxic metabolites are easily generated by traditional strains. CotA laccase and peroxide reductase Prx secreted by the strain can efficiently degrade ZEN and derivatives alpha-zearalenol and beta-zearalenol, the products are low-toxicity C17H24O4 and C12H16O4, and harmful by-products such as alpha-ZEL / beta-ZEL and the like do not exist. The strain and recombinase thereof are suitable for a high-temperature alkaline environment for feed processing, and can be applied to biological detoxification of ZEN pollution in food and feed on a large scale through an immobilized carrier.

The invention relates to the related technical field of molecular biology and cell engineering, in particular to an FLP-IN-PIEC cell line and a construction method and application thereof, and the construction method comprises the following steps: step 1, constructing CRISPR / Cas9 site-specific cleavage plasmid containing a pig HIPP11 safety site sequence; step 2, constructing a donor plasmid Donor-FRT containing a homologous arm of a safety site of the HIPP11; 3, jointly transfecting plasmids in the step 1 and the step 2 into a porcine hip artery endothelial cell line (PIEC) to obtain an FLP-IN-PIEC cell line for expressing red fluorescence; and 4, under the action of Flp recombinase, transfecting an exogenous gene donor plasmid containing an FRT site to the FLP-IN-PIEC cell line obtained in the step 3, and driving the expression of a target gene through an optimized strong promoter to realize fixed-point transfer and efficient expression of the exogenous gene in the pig (PIEC) cell line. Different from an existing Flp-In system depending on a random integration strategy, the construction method disclosed by the invention emphasizes fixed-point integration and promoter optimization, and improves the homogeneity, stability and screening efficiency of exogenous gene expression.

The present invention pertains to a method for generating a spacer-specific DNA recombining enzyme (DRE), the method comprising the steps of: a) providing a library of expression vectors encoding a plurality of variants of a DRE (vDRE), wherein the amino acid sequences of the vDREs comprise one or more amino acid modifications in comparison to a non-variant DRE (nvDRE) from which the vDREs are derived, wherein the nvDRE binds to a first target site comprising in 5′ to 3′ direction a half-site A (HSA), a spacer (Spacer) and a half-site B (HSB), wherein each expression vector comprises: (i) a first region comprising a nucleotide sequence encoding one of the vDREs from among the plurality of vDREs operably linked to an expression control sequence, and (ii) a second region comprising a nucleotide sequence comprising in 5′ to 3′ direction a first target site, an insert nucleotide sequence (INS) of a length at least 1 nucleotide and a second target site, wherein each of the first and second target site comprises the HSA, a variant spacer (vSpacer) and the HSB, wherein the vSpacer differs by at least one nucleotide from the Spacer; b) introducing the library of expression vectors into host cells; c) expressing the plurality of vDREs in the host cells; d) optionally isolating DNA from the host cells; and e) determining whether the vDRE shows activity on the target sites of at least one expression vector.

The invention discloses recombinant poria cocos chitin endonuclease as well as a gene, a preparation method and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO.1 in a sequence table. According to the nucleotide sequence as shown in SEQ ID NO.1, high-level recombinant secretory expression of target protein in pichia pastoris host bacteria can be realized by taking pichia pastoris inducible expression plasmid pPICZ alpha A as a carrier. Under a shake flask fermentation condition, the average secretory expression quantity can reach 300.3 mg / L. The recombinant enzyme obtained by nickel ion affinity chromatography purification not only can effectively hydrolyze colloidal chitin to generate chitosan oligosaccharide (COS), but also can directly degrade chitin-containing raw materials such as shrimps, crabs and the like to generate COS. In addition, the enzyme also shows the activity of hydrolyzing polysaccharides such as cellulose, pectin, agar and the like, and has important potential values in the fields of functional chitosan oligosaccharide preparation, biological energy production, medicine development, feed, environmental protection, agricultural application and the like.

The invention discloses a kit and a device for detecting acinetobacter baumannii. The kit comprises a first primer, a second primer, recombinase polymerase, Cas12a protein, crRNA and ssDNA, wherein the sequence of the first primer is as shown in SEQ ID NO.1; the sequence of the second primer is as shown in SEQ ID NO. 2; the sequence of the crRNA is as shown in SEQ ID NO.3, the crRNA is modified with a light-sensitive chemical group for preventing the combination of the crRNA and target nucleic acid, and the light-sensitive chemical group falls off from the crRNA when irradiated by light with a predetermined wavelength; the sequence of the ssDNA is as shown in SEQ ID NO.4, one end of the ssDNA is modified with a fluorophore, and the other end of the ssDNA is modified with a quenching group. By adopting the kit or the device provided by the invention, the acinetobacter baumannii can be accurately detected, the working efficiency can be improved, and the risk of cross contamination can be reduced.

The purpose of the present invention is to provide a novel method for targeting a neuron population that is a neural circuit element which has more limited functional localization. The purpose of the present invention is also to provide a method for treating a disease or the like by targeting a specific neuron population involved in a disease or the like by said novel method and controlling the neural activity thereof.　The present invention is a vector set used for targeting a specific neuron population in a neural circuit, said vector set being characterized by comprising first to third vectors, wherein: the first vector is an antegrade trans-synaptic virus vector that contains a base sequence which codes for a split-Cre recombinase consisting of a C-terminal-side sequence of a Cre recombinase; the second vector is a retrograde trans-synaptic virus vector that contains a base sequence which codes for a split-Cre recombinase consisting of an N-terminal-side sequence of a Cre recombinase; the third vector is a non-trans-synaptic virus vector that contains an inverted sequence of a gene sequence which codes for a desired protein and that has a loxP sequence and a loxP mutation sequence which face opposite to each other and which are at both ends of the inverted sequence; and a functional Cre recombinase is formed in a cell that has been infected with the first vector and the second vector.

The construct for site-specific insertion of DNA fragments and transgenes into the genome comprises a recombinant enzyme. This enzyme contains a modified HIV-1 integrase (HIV IN) tetramer with multiple mutations favoring a heterodimer structure in which the first and second outer monomers are bound to the first and second inner monomers, respectively. A transcription activator-like effector (TALE) is bound to the N-terminal region of the outer monomer via a polypeptide linker. The two TALEs confer sequence specificity to the enzyme and prevent non-specific interactions with the target DNA structure. The modified HIV IN monomer ensures that the TALE binds to the outermost monomer and that the TALE is positioned relative to the catalytic region of HIV IN. A polynucleotide is bound to the inner monomer of HIV IN. When the TALE is transported to the nucleus, either alone or within a lentiviral capsid, it interacts with the DNA to localize HIV IN, which then incorporates the polynucleotide.