Evidence map›Paper›PMID 24959875›Full record

ArticlePloS one2014

HomeRun Vector Assembly System: a flexible and standardized cloning system for assembly of multi-modular DNA constructs.

Ming V Li, Dip Shukla, Brian H Rhodes, Anjali Lall, Jingmin Shu, Branden S Moriarity, David A Largaespada

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

6 citing papers in PubMed.

  1. Review
  2. Article
  3. Review
  4. Article
  5. Article
  6. Bricks and blueprints: methods and standards for DNA assembly.Nature reviews. Molecular cell biology · 2015
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Ming V LiDivision of Endocrinology and Diabetes, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
Dip ShuklaDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, United States of America.
Brian H RhodesDepartment of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, United States of America.
Anjali LallDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, United States of America.
Jingmin ShuDivision of Endocrinology and Diabetes, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America; Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, United States of America; Center for Genome Engineering and Institute of Human Genetics, University of Minnesota, Minneapolis, Minnesota, United States of America; Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, United States of America.
Branden S MoriarityDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, United States of America; Center for Genome Engineering and Institute of Human Genetics, University of Minnesota, Minneapolis, Minnesota, United States of America; Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, United States of America.
David A LargaespadaDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, United States of America; Center for Genome Engineering and Institute of Human Genetics, University of Minnesota, Minneapolis, Minnesota, United States of America; Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, United States of America.

Funding

TRAINING GRANT IN DIABETES, ENDOCRINOLOGY, &METABOLISMT32DK007203 · NIDDK · UNIVERSITY OF MINNESOTA TWIN CITIES · PI LISA Senye CHOW, Douglas G Mashek · 1986 to 2026
$4.3M
NIDDK NIH HHS 5 T32 DK007203-35NIDDK NIH HHS T32 DK007203
6 · The paper itself

Abstract

Advances in molecular and synthetic biology call for efficient assembly of multi-modular DNA constructs. We hereby present a novel modular cloning method that obviates the need for restriction endonucleases and significantly improves the efficiency in the design and construction of complex DNA molecules by standardizing all DNA elements and cloning reactions. Our system, named HomeRun Vector Assembly System (HVAS), employs a three-tiered vector series that utilizes both multisite gateway cloning and homing endonucleases, with the former building individual functional modules and the latter linking modules into the final construct. As a proof-of-principle, we first built a two-module construct that supported doxycycline-induced expression of green fluorescent protein (GFP). Further, with a three-module construct we showed quantitatively that there was minimal promoter leakage between neighbouring modules. Finally, we developed a method, in vitro Cre recombinase-mediated cassette exchange (RMCE) cloning, to regenerate a gateway destination vector from a previous multisite gateway cloning reaction, allowing access to existing DNA element libraries in conventional gateway entry clones, and simple creation of constructs ready for in vivo RMCE. We believe these methods constitute a useful addition to the standard molecular cloning techniques that could potentially support industrial scale synthesis of DNA constructs.

Indexed as

Cloning, MolecularDNAGenetic VectorsHCT116 CellsHumansDNA

Identifiers

PMID24959875
PMCPMC4069157

What OpenQuestion holds

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LicenceCC BY
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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.