Enhanced gene disruption by programmable nucleases delivered by a minicircle vector.

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  • Additional Information
    • Publication Year:
      2014
    • NAICS/Industry Codes:
      NAICS/Industry Codes 541711 Research and Development in Biotechnology
    • Abstract:
      Targeted genetic modification using programmable nucleases such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) is of great value in biomedical research, medicine and biotechnology. Minicircle vectors, which lack extraneous bacterial sequences, have several advantages over conventional plasmids for transgene delivery. Here, for the first time, we delivered programmable nucleases into human cells using transient transfection of a minicircle vector and compared the results with those obtained using a conventional plasmid. Surrogate reporter assays and T7 endonuclease analyses revealed that cells in the minicircle vector group displayed significantly higher mutation frequencies at the target sites than those in the conventional plasmid group. Quantitative PCR and reverse transcription-PCR showed higher vector copy number and programmable nuclease transcript levels, respectively, in 293T cells after minicircle versus conventional plasmid vector transfection. In addition, tryphan blue staining and flow cytometry after annexin V and propidium iodide staining showed that cell viability was also significantly higher in the minicircle group than in the conventional plasmid group. Taken together, our results show that gene disruption using minicircle vector-mediated delivery of ZFNs and TALENs is a more efficient, safer and less toxic method than using a conventional plasmid, and indicate that the minicircle vector could serve as an advanced delivery method for programmable nucleases. [ABSTRACT FROM AUTHOR]
    • Abstract:
      Copyright of Gene Therapy is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
    • ISSN:
      09697128
    • Accession Number:
      99277308
  • Citations
    • ABNT:
      DAD, A.-B. et al. Enhanced gene disruption by programmable nucleases delivered by a minicircle vector. Gene Therapy, [s. l.], v. 21, n. 11, p. 921, 2014. Disponível em: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308. Acesso em: 9 dez. 2019.
    • AMA:
      Dad A-B, Ramakrishna S, Song M, Kim H. Enhanced gene disruption by programmable nucleases delivered by a minicircle vector. Gene Therapy. 2014;21(11):921. http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308. Accessed December 9, 2019.
    • APA:
      Dad, A.-B., Ramakrishna, S., Song, M., & Kim, H. (2014). Enhanced gene disruption by programmable nucleases delivered by a minicircle vector. Gene Therapy, 21(11), 921. Retrieved from http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308
    • Chicago/Turabian: Author-Date:
      Dad, A-BK, S Ramakrishna, M Song, and H Kim. 2014. “Enhanced Gene Disruption by Programmable Nucleases Delivered by a Minicircle Vector.” Gene Therapy 21 (11): 921. http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308.
    • Harvard:
      Dad, A.-B. et al. (2014) ‘Enhanced gene disruption by programmable nucleases delivered by a minicircle vector’, Gene Therapy, 21(11), p. 921. Available at: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308 (Accessed: 9 December 2019).
    • Harvard: Australian:
      Dad, A-B, Ramakrishna, S, Song, M & Kim, H 2014, ‘Enhanced gene disruption by programmable nucleases delivered by a minicircle vector’, Gene Therapy, vol. 21, no. 11, p. 921, viewed 9 December 2019, .
    • MLA:
      Dad, A.BK, et al. “Enhanced Gene Disruption by Programmable Nucleases Delivered by a Minicircle Vector.” Gene Therapy, vol. 21, no. 11, Nov. 2014, p. 921. EBSCOhost, search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308.
    • Chicago/Turabian: Humanities:
      Dad, A-BK, S Ramakrishna, M Song, and H Kim. “Enhanced Gene Disruption by Programmable Nucleases Delivered by a Minicircle Vector.” Gene Therapy 21, no. 11 (November 2014): 921. http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308.
    • Vancouver/ICMJE:
      Dad A-B, Ramakrishna S, Song M, Kim H. Enhanced gene disruption by programmable nucleases delivered by a minicircle vector. Gene Therapy [Internet]. 2014 Nov [cited 2019 Dec 9];21(11):921. Available from: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=edb&AN=99277308