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1999 5
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95 results

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T5 exonuclease-dependent assembly offers a low-cost method for efficient cloning and site-directed mutagenesis.
Xia Y, Li K, Li J, Wang T, Gu L, Xun L. Xia Y, et al. Nucleic Acids Res. 2019 Feb 20;47(3):e15. doi: 10.1093/nar/gky1169. Nucleic Acids Res. 2019. PMID: 30462336 Free PMC article.
Here, we present a 'T5 exonuclease DNA assembly' (TEDA) method that only uses a 5'-3' exonuclease. DNA fragments with short homologous ends were treated by T5 exonuclease and then transformed into Escherichia coli to produce clone colonies. ...
Here, we present a 'T5 exonuclease DNA assembly' (TEDA) method that only uses a 5'-3' exonuclease. DNA fragments with s …
Addition of the T5 exonuclease increases the prime editing efficiency in plants.
Liang Z, Wu Y, Guo Y, Wei S. Liang Z, et al. J Genet Genomics. 2023 Aug;50(8):582-588. doi: 10.1016/j.jgg.2023.03.008. Epub 2023 Mar 21. J Genet Genomics. 2023. PMID: 36958601 Free article.
Here we develop different versions of PE2 by adding the 5'-to-3' exonuclease at different positions of the nCas9-M-MLV RT fusion protein. PE2 (v2), in which the T5 exonuclease fused to the N-terminus of the nCas9-MMLV fusion protein enhances prime editing eff …
Here we develop different versions of PE2 by adding the 5'-to-3' exonuclease at different positions of the nCas9-M-MLV RT fusion prot …
TLTC, a T5 exonuclease-mediated low-temperature DNA cloning method.
Yu F, Li X, Wang F, Liu Y, Zhai C, Li W, Ma L, Chen W. Yu F, et al. Front Bioeng Biotechnol. 2023 Aug 11;11:1167534. doi: 10.3389/fbioe.2023.1167534. eCollection 2023. Front Bioeng Biotechnol. 2023. PMID: 37635997 Free PMC article.
Here, we developed a rapid and efficient DNA-assembling method for routine laboratory work. We discovered that the cleavage speed of T5 exonuclease is approximately 3 nt/min at 0C and hence developed a T5 exonuclease-mediated low-temperature sequence- …
Here, we developed a rapid and efficient DNA-assembling method for routine laboratory work. We discovered that the cleavage speed of T5 …
Enzymatic assembly of overlapping DNA fragments.
Gibson DG. Gibson DG. Methods Enzymol. 2011;498:349-61. doi: 10.1016/B978-0-12-385120-8.00015-2. Methods Enzymol. 2011. PMID: 21601685 Free PMC article.
The second method uses 3'-exonuclease III (ExoIII), antibody-bound Taq pol, and Taq lig in a one-step thermocycled reaction. The third method employs 5'-T5 exonuclease, Phusion DNA polymerase, and Taq lig in a one-step isothermal reaction and can be used to a …
The second method uses 3'-exonuclease III (ExoIII), antibody-bound Taq pol, and Taq lig in a one-step thermocycled reaction. The thir …
A T5 Exonuclease-Based Assay for DNA Topoisomerases and DNA Intercalators.
Deng Z, Leng F. Deng Z, et al. ACS Omega. 2021 Apr 28;6(18):12205-12212. doi: 10.1021/acsomega.1c00962. eCollection 2021 May 11. ACS Omega. 2021. PMID: 34056374 Free PMC article.
We report here the establishment of a fluorescence-based, low-cost HTS assay to identify topoisomerase inhibitors. This HTS assay is based on a unique property of T5 exonuclease that can completely digest supercoiled plasmid pAB1 containing an "AT" hairpin structure …
We report here the establishment of a fluorescence-based, low-cost HTS assay to identify topoisomerase inhibitors. This HTS assay is based o …
Development of miniature base editors using engineered IscB nickase.
Han D, Xiao Q, Wang Y, Zhang H, Dong X, Li G, Kong X, Wang S, Song J, Zhang W, Zhou J, Bi L, Yuan Y, Shi L, Zhong N, Yang H, Zhou Y. Han D, et al. Nat Methods. 2023 Jul;20(7):1029-1036. doi: 10.1038/s41592-023-01898-9. Epub 2023 May 25. Nat Methods. 2023. PMID: 37231266
Here we describe the engineering of OgeuIscB and its corresponding omegaRNA to develop an IscB system that is highly efficient in mammalian systems, named enIscB. By fusing enIscB with T5 exonuclease (T5E), we found enIscB-T5E exhibited comparable targeting efficien …
Here we describe the engineering of OgeuIscB and its corresponding omegaRNA to develop an IscB system that is highly efficient in mammalian …
A Fluorescence-Based, T5 Exonuclease-Amplified DNA Cleavage Assay for Discovering Bacterial DNA Gyrase Poisons.
Dias M, Chapagain T, Leng F. Dias M, et al. bioRxiv [Preprint]. 2023 Oct 16:2023.10.16.562555. doi: 10.1101/2023.10.16.562555. bioRxiv. 2023. PMID: 37904923 Free PMC article. Preprint.
Proteinase K digestion results in producing small DNA fragments. T5 exonuclease, selectively digesting linear and nicked DNA, can fully digest the fragmented linear DNA molecules and, thus, "amplify" the decrease in fluorescence signal of the DNA cleavage products a …
Proteinase K digestion results in producing small DNA fragments. T5 exonuclease, selectively digesting linear and nicked DNA, …
Fusing T5 exonuclease with Cas9 and Cas12a increases the frequency and size of deletion at target sites.
Zhang Q, Yin K, Liu G, Li S, Li M, Qiu JL. Zhang Q, et al. Sci China Life Sci. 2020 Dec;63(12):1918-1927. doi: 10.1007/s11427-020-1671-6. Epub 2020 May 6. Sci China Life Sci. 2020. PMID: 32382982
Here we show that it is possible to create such deletions with a single guide RNA by fusing Cas9 or Cas12a with T5 exonuclease (T5exo). These fusion constructs were found to increase both the frequency and size of deletions at target loci in rice protoplasts and see …
Here we show that it is possible to create such deletions with a single guide RNA by fusing Cas9 or Cas12a with T5 exonuclease …
T5 Exonuclease Hydrolysis of Hepatitis B Virus Replicative Intermediates Allows Reliable Quantification and Fast Drug Efficacy Testing of Covalently Closed Circular DNA by PCR.
Qu B, Ni Y, Lempp FA, Vondran FWR, Urban S. Qu B, et al. J Virol. 2018 Nov 12;92(23):e01117-18. doi: 10.1128/JVI.01117-18. Print 2018 Dec 1. J Virol. 2018. PMID: 30232183 Free PMC article.
However, PCR-based assays for cccDNA quantification show a principally constrained specificity when high levels of input virus or replicative intermediates are present. Here, we characterized T5 exonuclease as a suitable enzyme for medium-throughput in vitro assays …
However, PCR-based assays for cccDNA quantification show a principally constrained specificity when high levels of input virus or replicativ …
Quantification of the hepatitis B virus cccDNA: evidence-based guidelines for monitoring the key obstacle of HBV cure.
Allweiss L, Testoni B, Yu M, Lucifora J, Ko C, Qu B, Lütgehetmann M, Guo H, Urban S, Fletcher SP, Protzer U, Levrero M, Zoulim F, Dandri M. Allweiss L, et al. Gut. 2023 May;72(5):972-983. doi: 10.1136/gutjnl-2022-328380. Epub 2023 Jan 27. Gut. 2023. PMID: 36707234 Free PMC article.
Each group compared different DNA extraction methods (Hirt extraction, total DNA extraction with or without proteinase K treatment (+PK/-PK)) and nuclease digestion protocols (plasmid-safe ATP-dependent DNase (PSD), T5 exonuclease, exonucleases I/III). Sample …
Each group compared different DNA extraction methods (Hirt extraction, total DNA extraction with or without proteinase K treatment (+PK/-PK) …
95 results