Literature DB >> 28426753

Correction: A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.

Daniel K Fowler, Scott Stewart, Steve Seredick, Judith S Eisen, Kryn Stankunas, Philip Washbourne.   

Abstract

[This corrects the article DOI: 10.1371/journal.pone.0159277.].

Entities:  

Year:  2017        PMID: 28426753      PMCID: PMC5398664          DOI: 10.1371/journal.pone.0176543

Source DB:  PubMed          Journal:  PLoS One        ISSN: 1932-6203            Impact factor:   3.240


In Fig 1, the orientation of vector-specific 5’ and 3’ sequences for pEpic and pEpic_Lite and derivatives of these vectors are incorrect. The correct configurations are all sense-strand oriented components. Please see the corrected Fig 1 here.
Fig 1

Overview of three-fragment MultiSite Gateway cloning and novel lentiviral destination vectors.

(A) Schematic of an LR recombination reaction and the resulting vector. Site-specific recombination events (red lines) between attR and attL sites from a 5’, middle, and 3’ entry vector with a destination vector replaces the ccdB/CmR selection cassette of the destination vector with the mobile DNA elements from the entry vectors, leaving destination vector-specific 5’ and 3’ sequences intact. (B) Schematic of lentiviral destination vectors pEpic and pEpic_Lite. attR3 and 4 sites flanking the ccdB/CmR selection cassette are positioned in an anti-sense orientation to viral RNA expression driven by a Rous sarcoma virus (RSV) promoter. pEpic_Lite lacks puromycin resistance (PuroR). LTR = long terminal repeat; RRE = Rev response element; cPPT = central polypurine tract; ccdB = E. coli ccdB toxin; CmR = chloramphenicol resistance; mPGK = mouse phosphoglycerate kinase promoter; WPRE = woodchuck hepatitis virus posttranslational regulatory element.

Overview of three-fragment MultiSite Gateway cloning and novel lentiviral destination vectors.

(A) Schematic of an LR recombination reaction and the resulting vector. Site-specific recombination events (red lines) between attR and attL sites from a 5’, middle, and 3’ entry vector with a destination vector replaces the ccdB/CmR selection cassette of the destination vector with the mobile DNA elements from the entry vectors, leaving destination vector-specific 5’ and 3’ sequences intact. (B) Schematic of lentiviral destination vectors pEpic and pEpic_Lite. attR3 and 4 sites flanking the ccdB/CmR selection cassette are positioned in an anti-sense orientation to viral RNA expression driven by a Rous sarcoma virus (RSV) promoter. pEpic_Lite lacks puromycin resistance (PuroR). LTR = long terminal repeat; RRE = Rev response element; cPPT = central polypurine tract; ccdB = E. coli ccdB toxin; CmR = chloramphenicol resistance; mPGK = mouse phosphoglycerate kinase promoter; WPRE = woodchuck hepatitis virus posttranslational regulatory element. The correct plasmids and their sequences have been deposited with Addgene (www.addgene.org), with the following catalog numbers: pEpic #84372; pEpic_Lite #84373.
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1.  A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.

Authors:  Daniel K Fowler; Scott Stewart; Steve Seredick; Judith S Eisen; Kryn Stankunas; Philip Washbourne
Journal:  PLoS One       Date:  2016-08-08       Impact factor: 3.240

  1 in total
  1 in total

1.  Neural stem cell quiescence and stemness are molecularly distinct outputs of the Notch3 signalling cascade in the vertebrate adult brain.

Authors:  Emmanuel Than-Trong; Sara Ortica-Gatti; Sébastien Mella; Chirag Nepal; Alessandro Alunni; Laure Bally-Cuif
Journal:  Development       Date:  2018-05-15       Impact factor: 6.868

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