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TALENs: a widely applicable technology for targeted genome editing
Open accessAbstract
Engineered nucleases enable the targeted alteration of nearly any gene in a wide range of cell types and organisms. The newly-developed transcription activator-like effector nucleases (TALENs) comprise a nonspecific DNA-cleaving nuclease fused to a DNA-binding domain that can be easily engineered so that TALENs can target essentially any sequence. The capability to quickly and efficiently alter genes using TALENs promises to have profound impacts on biological research and to yield potential therapeutic strategies for genetic diseases.
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“Each repeat binds to a single nucleotide, and the amino acids residues in the positions 12 and 13 of that repeat determine which nucleotide is bound [91].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [91]
In the citing paper
“The discovery that TALEs use a simple modular code for DNA recognition has provided an alternative platform for genome editing and gene disruption.”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [66]
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“21 The TALE repeat code was also confirmed by another group by performing a computational analysis of the binding specificities of naturally occurring TALEs 22 .”
Genome editing with engineered zinc finger nucleases
Urnov, F. D., Rebar, E. J., Holmes, M. C., Zhang, H. S., & Gregory, P. D. (2010) · Nature Reviews Genetics · doi:10.1038/nrg2842 · Cited by 2,285
A TALE nuclease architecture for efficient genome editing
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“Alternatively, precise insertions have also been introduced into endogenous human genes using HDR with TALENs and a double-stranded homologous donor template plasmid 19, 20 .”
InfluentialA Simple Cipher Governs DNA Recognition by TAL Effectors
Moscou, M., & Bogdanove, A. J. (2009) · Science · doi:10.1126/science.1178817 · Cited by 2,050 · cited 6×
“4 DNA binding by these TALEs is mediated by arrays of highly conserved 33–35 amino acid repeats flanked by additional TALE-derived domains at the amino-and carboxy-terminal ends of the array (Figure 1c).”
InfluentialČermák, T., Doyle, E., Christian, M. L., Wang, L., Zhang, Y., Schmidt, C. L., Baller, J. A., Somia, N. V., Bogdanove, A. J., & Voytas, D. F. (2011) · Nucleic Acids Research · doi:10.1093/nar/gkr218 · Cited by 2,034 · cited 6×
“To date, TALENs have been used to make knockout mutations in Arabidopsis 15 and to introduce resistance to infection by Xanthomonas bacteria in rice by disrupting the target sites of naturally occurring TALEs that contribute to pathogenicity 16 .”
InfluentialHybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain.
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“24 The TALEN framework was then refined by showing that nuclease activities could be greatly enhanced by truncating the length of this carboxy-terminal TALE-derived sequence 19 , 68 .”
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“…38 , catfish 39 , sea urchin 40 , frog 41 , pig 42 , cattle 43 , cricket 8 , rabbit 44 , silkworm 45 , butterfly 46 , mouse 47, 48 , rat 49, 50 , soybean 51, 52 , thale cress 53, 54 , corn 55 , tobacco 56 , petunia 57 , hamster cells 58 , and human somatic 59, 60 and pluripotent stem cells 61–63 .”
Vascular Endothelial Growth Factor
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“Although the high success rate and robust targeting range of dimeric TALENs has been recently established, 18 relatively less is known about these parameters for monomeric engineered TALE-based transcriptional activators or repressors.”
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“Traditional gene editing tools principally include Zinc Finger Nucleases (ZFNs) [30–32], Transcriptional Activator Effector Nucleases (TALENs) [33], and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) [34].”
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“Additional tools for genome editing in eukaryotes include engineered zinc-finger nucleases (ZFNs, [20, 21]) and transcription activator-like effector nucleases (TALENs, [20, 22, 23]).”
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“…(Carroll et al., 2008; Cui et al., 2011; Urnov et al., 2010), transcription activator-like effector nucleases (TALENs) (Bogdanove and Voytas, 2011; Joung and Sander, 2013; Sommer et al., 2014) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas)…”
BackgroundCRISPR ‐Cas9 technology and its application in haematological disorders
Zhang, H., & McCarty, N. (2016) · British Journal of Haematology · doi:10.1111/bjh.14297 · Cited by 37
“Although ZFNs and TALENs have resulted in more efficient and applicable site-specific genome engineering (Joung & Sander, 2013; Urnov, 2014), the expansion of their applications has also been hampered by complex procedures, low specificity and high cost (Peng et al, 2015; Sanchez-Rivera & Jacks,…”
BackgroundGlover, B., Akinbo, O., Savadogo, M., Timpo, S. E., Lemgo, G., Sinebo, W., Akile, S., Obukosia, S. D., Ouedraogo, J. T., Ndomondo-Sigonda, M., et al. (2018) · BMC Proceedings · doi:10.1186/s12919-018-0108-y · Cited by 36
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“Based on the presence or absence of donor DNA template the stimulated double-stranded DNA breaks can be repaired, respectively, by homology-directed repair (HDR) or non-homologous end joining (NHEJ) pathways [25,26].”
BackgroundComputational Tools for Investigating RNA-Protein Interaction Partners
Muppirala, U. K. (2013) · Journal of Computer Science & Systems Biology · doi:10.4172/jcsb.1000115 · Cited by 36
“Engineered DNA binding proteins, such as ZFNs and TALENS, have become enormously powerful tools for genome engineering, and are poised to enter clinical settings [56-58].”
BackgroundTranslating Lung Function Genome-Wide Association Study (GWAS) Findings
Kheirallah, A. K., Miller, S., Hall, I. P., & Sayers, I. (2016) · Advances in genetics · doi:10.1016/bs.adgen.2015.12.002 · Cited by 36
“Then non-homologous end-joining or homologydirected repair are activated, thus allowing editing of target sites (Joung & Sander, 2013).”
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“Recently, TALEN has emerged as an efficient tool for creating genetically engineered models in various species via genome editing and the introduction of targeted double-strand breaks (DSBs).(42) This method circumvents the requirement for embryonic stem cells and opens up the application of gene targeting for many sp…”
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Functional interrogation of non-coding DNA through CRISPR genome editing
Canver, M. C., Bauer, D. E., & Orkin, S. H. (2017) · Methods · doi:10.1016/j.ymeth.2017.03.008 · Cited by 36
“The development of genome editing technologies including zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) offered new methods to identify functional non-coding sequences through loss- and gain-of-function genetic studies [10,11].”
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“Advances in genome engineering (2, 94-96) allow researchers to study the impact of genetic variants of interest, or to manipulate specific transcripts or epigenetic factors, on an isogenic background, thereby removing variance due to genetic differences between individuals (Figure 2; although the impact of variants sh…”
Clinical genome editing to treat sickle cell disease—A brief update
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“Compared to ZFNs, TALEN is significantly simpler to construct, has the ability to be synthesized to target a specific place in the genome, and is not constrained by the length of the sequence it can bind to (Joung and Sander 2013).”
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“35 Base editors represent a groundbreaking advancement in genome editing technologies, offering the ability to introduce precise single-nucleotide substitutions without inducing DSBs or relying on donor DNA templates.”
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“Prior to CRISPR, targeted endonucleases such as zinc-finger nucleases (ZFNs) 9 and transcription activator-like effector nucleases (TALENs) 10 required laborious re-engineering for each new genomic site.”
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“Chimeric enzymes, such as zinc-finger nucleases (ZFNs) [20] and transcription activator-like effector nucleases (TALENs) [21], harbor both a domain specifying the target sequence and an endonuclease domain to affect a DSB at the desired site in the genome.”
BackgroundCohn, M. M., Morbitzer, R., Lahaye, T., & Staskawicz, B. J. (2015) · Molecular Plant Pathology · doi:10.1111/mpp.12337 · Cited by 33
“The pepper (Capsicum annuum) transcription factor UPA20, a tar- get of the TAL effector AvrBs3 from Xanthomonas euvesicatoria (Xe), contributes to symptom formation by inducing cell hypertro- phy (Kay et al., 2007).”
BackgroundAdvancing biotechnology with CRISPR/Cas9: recent applications and patent landscape
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“Only recently discovered, CRISPR/Cas9 technology has already been enhanced to the point of fulfilling most of the genome editing and gene regulation currently demanded, ranging from the ability to perform multiple gene insertions, gene knockouts, combinatorial libraries, to advanced fine-tuning of biosynthetic pathway…”
BackgroundPRISM offers a comprehensive genomic approach to transcription factor function predictionPreprint
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“At the same time, when Hendel’s group tried to insert shorter sequence with 400 bp homology arms into the IL2RG locus in K562 cells using TALEN nuclease, the knock-in efficiency was the same (17”
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“TALEN, the second-generation genome engineering effector, was also designed to function as a dimer by connecting a TAL effector-like motif to FokI nuclease domain (Joung and Sander, 2013)(Fig.”
BackgroundApplications and Major Achievements of Genome Editing in Vegetable Crops: A Review
Kim, Y., Kang, Y., Yang, E.-Y., Cho, M.-C., Schafleitner, R., Lee, J. H., & Jang, S. (2021) · Frontiers in Plant Science · doi:10.3389/fpls.2021.688980 · Cited by 33
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“A 1.1-kb human elongation factor 1α (EF1α) promoter was then amplified from pFB-EF1α-EGFP-hyg-lox [11] and cloned into the above plasmid vector using BamHI/NotI to drive the expression of ZFNs.”
BackgroundInfluentialScreening Methods to Identify TALEN-Mediated Knockout Mice
Nakagawa, Y., Yamamoto, T., Suzuki, K., Araki, K., Takeda, N., Ohmuraya, M., & Sakuma, T. (2014) · EXPERIMENTAL ANIMALS · doi:10.1538/expanim.63.79 · Cited by 31
Translating human genetics into mouse: The impact of ultra‐rapid in vivo genome editing
Aida, T., Imahashi, R., & Tanaka, K. (2013) · Development Growth & Differentiation · doi:10.1111/dgd.12101 · Cited by 33
“1, Urnov et al. 2010; Joung & Sander 2013).”
Ferguson, C., McKay, M., Harris, R. A., & Homanics, G. E. (2013) · Alcohol · doi:10.1016/j.alcohol.2013.09.043 · Cited by 31
“TALEN proteins are customizable, engineered nucleases that can be targeted to virtually any DNA sequence in the genome (for reviews, see Bogdanove & Voytas, 2011; Joung & Sander, 2013).”
BackgroundMethodProgress in gene therapy for primary immunodeficiencies using lentiviral vectors
Sauer, A. V., Lorenzo, B. D., Carriglio, N., & Aiuti, A. (2014) · Current Opinion in Allergy and Clinical Immunology · doi:10.1097/aci.0000000000000114 · Cited by 32
“Employed artificial endonucleases include zinc finger nucleases [87], transcription activator-like effector nucleases [88] and RNA-guided nucleases [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated gene (Cas)] [89].”
BackgroundThe mitochondrial transporter SLC25A25 links ciliary TRPP2 signaling and cellular metabolism
Hofherr, A., Seger, C., Fitzpatrick, F., Busch, T., Michel, E., Luan, J., Osterried, L., Linden, F., Kramer-Zucker, A. G., Wakimoto, B. T., et al. (2018) · PLoS Biology · doi:10.1371/journal.pbio.2005651 · Cited by 32
“Two TALEN pairs per gene were designed to induce parallel 5 and 3 genomic DNA double-strand breaks facilitating the deletion of the respective gene by nonhomologous endjoining [64].”
Illuminating the genome-wide activity of genome editors for safe and effective therapeutics
Cheng, Y., & Tsai, S. Q. (2018) · Genome biology · doi:10.1186/s13059-018-1610-2 · Cited by 32
Histone modifications, DNA methylation, and the epigenetic code of alcohol use disorder
Bohnsack, J. P., & Pandey, S. C. (2020) · International review of neurobiology · doi:10.1016/bs.irn.2020.08.005 · Cited by 32
“Previously, this involved the use of engineered transcription activator-like effector nucleases (TALENs), which involved protein engineering and was costly and labor intensive (Joung and Sander, 2013).”
BackgroundGenome editing via non-viral delivery platforms: current progress in personalized cancer therapy
Lan, T., Que, H., Luo, M., Zhao, X., & Wei, X. (2022) · Molecular Cancer · doi:10.1186/s12943-022-01550-8 · Cited by 31
“Transcription activator‐like effector nucleases Transcription activator-like effector nuclease (TALEN) system is established by fusing transcription activatorlike effector (TALE) with DNA cleavage domain [72].”
BackgroundMethodReview: Precision Medicine Approaches for Genetic Cardiomyopathy: Targeting Phospholamban R14del
Deiman, F. E., Bömer, N., van der Meer, P., & Beverborg, N. G. (2022) · Current Heart Failure Reports · doi:10.1007/s11897-022-00558-x · Cited by 31
Genetic Advances in Microphthalmia
Plaisancié, J., Calvas, P., & Chassaing, N. (2016) · Journal of Pediatric Genetics · doi:10.1055/s-0036-1592350 · Cited by 31
Computational design of sequence-specific DNA-binding proteins
Glasscock, C. J., Pecoraro, R. J., McHugh, R., Doyle, L. A., Chen, W., Boivin, O., Lonnquist, B., Na, E., Politanska, Y., Haddox, H. K., et al. (2025) · Nature Structural & Molecular Biology · doi:10.1038/s41594-025-01669-4 · Cited by 58
Reichenbach, P., Attianese, G. M. P. G., Ouchen, K., Cribioli, E., Triboulet, M. M., Ash, S., Saillard, M., de Silly, R. V., Coukos, G., & Irving, M. B. (2023) · Nature Biomedical Engineering · doi:10.1038/s41551-023-01013-5 · Cited by 32
Sakane, Y., Sakuma, T., Kashiwagi, K., Kashiwagi, A., Yamamoto, T., & Suzuki, K. (2013) · Development Growth & Differentiation · doi:10.1111/dgd.12105 · Cited by 30
“TALEN architecture comprises tandem repeats of DNA-binding modules, N-terminal domain, C-terminal domain, and nuclease domain of FokI (Miller et al. 2011; Scholze & Boch 2011; Joung & Sander 2013).”
BackgroundCRISPR‐Mediated Gene Targeting of Human Induced Pluripotent Stem Cells
Byrne, S. M., & Church, G. M. (2015) · Current Protocols in Stem Cell Biology · doi:10.1002/9780470151808.sc05a08s35 · Cited by 30
“…activator-like effector nucleases (TALENs), or CRISPR/Cas9 nucleases have enormously expanded our ability to engineer genetic changes in human cells (Urnov et al., 2010; Joung and Sander, 2012; Mali et al., 2013b), particularly human iPSCs, which were previously resistant to gene editing.”
BackgroundEnergy biotechnology in the CRISPR-Cas9 era
Estrela, R., & Cate, J. H. D. (2016) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2016.01.005 · Cited by 30
Shen, Y., Xu, K., Yuan, Z., Guo, J., Zhao, H., Zhang, X., Zhao, L., Qing, Y., Li, H., Pan, W., et al. (2017) · Journal of Translational Medicine · doi:10.1186/s12967-017-1327-0 · Cited by 30
Sehgal, N., Sylves, M. E., Sahoo, A., Chow, J. L., Walker, S. E., Cullen, P. J., & Berry, J. O. (2018) · Biochemistry and Molecular Biology Education · doi:10.1002/bmb.21175 · Cited by 30
Genome-wide protein phylogenies for four African cichlid species
Varadarajan, A. R., Mopuri, R., Streelman, J. T., & McGrath, P. T. (2018) · BMC Evolutionary Biology · doi:10.1186/s12862-017-1072-2 · Cited by 30
“…sequencing costs and the development of broadly applicable genetic tools like TALENs and CRISPR/Cas9 has facilitated the development of a large number of new species as model organisms (Joung and Sander 2013, Doudna and Charpentier 2014, Hsu, Lander et al. 2014,Nemudryi, Valetdinova et al. 2014).”
Araldi, R. P., Khalil, C., Grignet, P. H., Teixeira, M. R., de Melo, T. C., Módolo, D. G., Fernandes, L. G. V., Ruiz, J. L. M., & de Souza, E. B. (2020) · Gene · doi:10.1016/j.gene.2020.144636 · Cited by 33
Wang, J., Huang, C., Guo, K., Ma, L., Meng, X., Wang, N., & Huo, Y. (2020) · Synthetic and Systems Biotechnology · doi:10.1016/j.synbio.2020.10.005 · Cited by 30
“Compared with 79 methods that utilize homing endonuclease I-SceI [29-31] and engineered endonucleases, 80 such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases 81 (TALENs) [32-35], to cleave dsDNA, clustered regularly interspaced short palindromic 82 repeats (CRISPR)/CRISPR-associate…”
MethodExperimental Models to Study Autism Spectrum Disorders: hiPSCs, Rodents and Zebrafish
Pensado‐López, A., Veiga-Rúa, S., Carracedo, Á., Allegue, C., & Sanchez, L. E. (2020) · Genes · doi:10.3390/genes11111376 · Cited by 34
“In comparison with ZFNs, TALENs are easier to synthesize, but the required protein design is still challenging [27,38].”
BackgroundVisualizing and Manipulating Temporal Signaling Dynamics with Fluorescence-Based Tools
Doupé, D. P., & Perrimon, N. (2014) · Science Signaling · doi:10.1126/scisignal.2005077 · Cited by 30
Gene editing to improve legume-rhizobia symbiosis in a changing climate
Jain, D., Jones, L., & Roy, S. (2022) · Current Opinion in Plant Biology · doi:10.1016/j.pbi.2022.102324 · Cited by 29
CRISPR/Cas-Mediated Genome Editing for Sugarcane Improvement
Mall, A. K., Manimekalai, R., Misra, V., Pandey, H., Srivastava, S., & Sharma, A. (2024) · Sugar Tech · doi:10.1007/s12355-023-01352-2 · Cited by 29
“Target sequences for TALEN pairs are typically 12–25 bp long (Joung and Sander 2013).”
Genome Manipulation Advances in Selected Aquaculture Organisms
Wang, J., Cheng, Y., Su, B., & Dunham, R. A. (2024) · Reviews in Aquaculture · doi:10.1111/raq.12988 · Cited by 33
Li, C., Qi, R., Singleterry, R., Hyle, J. W., Balch, A., Li, X., Sublett, J. E., Berns, H., Valentine, M. B., Valentine, V. A., et al. (2014) · Molecular and Cellular Biology · doi:10.1128/mcb.00023-14 · Cited by 29
SH2B1 is critical for the regulation of cardiac remodelling in response to pressure overload
Wu, G., Liu, Y., Huang, H., Tang, Y., Liu, W., Mei, Y., Wan, N., Liu, X., & Huang, C. (2015) · Cardiovascular Research · doi:10.1093/cvr/cvv170 · Cited by 29
“Importantly, genome-editing technologies such as designer TALENs permit targeted genome modifications in rats and can be designed to target any DNA sequence.(52) Thus, we successfully established a SH2B1-knockout rat model by utilizing TALENs in the SD rat background.”
MethodThe therapeutic landscape of HIV-1 via genome editing
Kwarteng, A., Ahuno, S. T., & Kwakye-Nuako, G. (2017) · AIDS Research and Therapy · doi:10.1186/s12981-017-0157-8 · Cited by 31
Advancements in CRISPR/Cas9 technology—Focusing on cancer therapeutics and beyond
Mirza, Z., & Karim, S. (2019) · Seminars in Cell and Developmental Biology · doi:10.1016/j.semcdb.2019.05.026 · Cited by 29
Application of CRISPR/Cas9 in Rapeseed for Gene Function Research and Genetic Improvement
Tian, Q. C., Li, B., Feng, Y., Zhao, W., Huang, J., & Chao, H. (2022) · Agronomy · doi:10.3390/agronomy12040824 · Cited by 33
Coengineering specificity, safety, and function into T cells for cancer immunotherapy
Attianese, G. M. P. G., Ash, S., & Irving, M. B. (2023) · Immunological Reviews · doi:10.1111/imr.13252 · Cited by 29
“47 Examples include transcription activator like effector nucleases, 48 zinc finger nucleases, 49 clustered regularly interspaced short palindromic repeats (CRISPR) 50,51 and viral vectors such as adenovirus, adeno-associated virus, 52 and γ - retrovirus and lentivirus.”
Background
Metadata
- Authors
- Joung, J. K., & Sander, J. D.
- Affiliations
- Harvard University (US)
- Corresponding author
- Joung, J. Keith
- Venue
- Nature Reviews Molecular Cell Biology
- Published
- 2012-11-21
- Volume
- 14
- Issue
- 1
- Pages
- 49-55
- ISSN
- 1471-0072 (ISSN-L)
- 1471-0080
- PMC
- PMC3547402
- OpenAlex
- W2061184966
- Cited by
- 1,650
- References
- 103
- Citations by year
- 2026: 65 · 2025: 106 · 2024: 128 · 2023: 119 · 2022: 123 · 2021: 102 · 2020: 112 · 2019: 122 · 2018: 114 · 2017: 103 · 2016: 114 · 2015: 159 · 2014: 175 · 2013: 107
- FWCI
- 12.87
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Nature Reviews Molecular Cell Biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Human Genome Research Institute (P50HG005550)
- National Cancer Institute (T32CA009216)
- National Institute of General Medical Sciences (DP1GM105378, R01GM088040)
- National Center for Chronic Disease Prevention and Health Promotion (DP1 GM105378)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- TALEN, targeted genome editing, engineered nucleases, genetic modification, genetic disease treatment
- MeSH terms
- Animals, Deoxyribonucleases, DNA, DNA-Binding Proteins, Humans, Protein Engineering, Transcriptional Activation, Genome, Protein Structure, Tertiary
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