Evidence map›Paper›PMID 28239692›Full record

ArticleNanoscale2017

High-throughput screening of microchip-synthesized genes in programmable double-emulsion droplets.

H F Chan, S Ma, J Tian, K W Leong

Open access · greenAbstract read
In one paragraph

Article in Nanoscale, 2017. 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
1.8field-weighted citation impact, top 16% of its field
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, 32 citations in OpenAlex.

  1. Article
  2. Review
  3. Article
  4. Article
  5. Article
  6. Article
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

4 authors at 1 institution in 2 countries.

H F ChanDepartment of Biomedical Engineering, Duke University, Durham, 27705, USA. kam.leong@columbia.edu and Department of Biomedical Engineering, Columbia University, New York, 10027, USA.
S MaDepartment of Biomedical Engineering, Duke University, Durham, 27705, USA. kam.leong@columbia.edu and General Biosystems, Inc. Morrisville, 27560 USA.
J TianDepartment of Biomedical Engineering, Duke University, Durham, 27705, USA. kam.leong@columbia.edu and General Biosystems, Inc. Morrisville, 27560 USA and Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
K W LeongDepartment of Biomedical Engineering, Duke University, Durham, 27705, USA. kam.leong@columbia.edu and Department of Biomedical Engineering, Columbia University, New York, 10027, USA and Department of Systems Biology, Columbia University, New York, 10027, USA.ORCID http://orcid.org/0000-0002-8133-4955
Duke University · US

Funding

Oral Therapy for Hemophilia AR01HL109442 · NHLBI · UNIVERSITY OF FLORIDA · PI DANIELL, HENRY, HERZOG, ROLAND W. · 2011 to 2015
$3.5M
Overcoming Immunosenescence by Nanoparticle-Mediated ActivationR01AI096305 · NIAID · DUKE UNIVERSITY · PI ABRAHAM, SOMAN N, LEONG, KAM W · 2012 to 2016
$1.8M
A synthetic biology approach to analyze evolution of programmed bacterial deathR01GM110494 · NIGMS · DUKE UNIVERSITY · PI YOU, LINGCHONG · 2014 to 2017
$1.2M
BD Biosciences InfluxS10OD020056 · OD · COLUMBIA UNIVERSITY HEALTH SCIENCES · PI SNOECK, HANS-WILLEM E · 2015 to 2015
$600k
NHLBI NIH HHS R01 HL109442NIAID NIH HHS R01 AI096305NIGMS NIH HHS R01 GM110494NIH HHS S10 OD020056
6 · The paper itself

Abstract

The rapid advances in synthetic biology and biotechnology are increasingly demanding high-throughput screening technology, such as screening of the functionalities of synthetic genes for optimization of protein expression. Compartmentalization of single cells in water-in-oil (W/O) emulsion droplets allows screening of a vast number of individualized assays, and recent advances in automated microfluidic devices further help realize the potential of droplet technology for high-throughput screening. However these single-emulsion droplets are incompatible with aqueous phase analysis and the inner droplet environment cannot easily communicate with the external phase. We present a high-throughput, miniaturized screening platform for microchip-synthesized genes using microfluidics-generated water-in-oil-in-water (W/O/W) double emulsion (DE) droplets that overcome these limitations. Synthetic gene variants of fluorescent proteins are synthesized with a custom-built microarray inkjet synthesizer, which are then screened for expression in Escherichia coli (E. coli) cells. Bacteria bearing individual fluorescent gene variants are encapsulated as single cells into DE droplets where fluorescence signals are enhanced by 100 times within 24 h of proliferation. Enrichment of functionally-correct genes by employing an error correction method is demonstrated by screening DE droplets containing fluorescent clones of bacteria with the red fluorescent protein (rfp) gene. Permeation of isopropyl β-d-1-thiogalactopyranoside (IPTG) through the thin oil layer from the external solution initiates target gene expression. The induced expression of the synthetic fluorescent proteins from at least ∼100 bacteria per droplet generates detectable fluorescence signals to enable fluorescence-activated cell sorting (FACS) of the intact droplets. This technology obviates time- and labor-intensive cell culture typically required in conventional bulk experiment.

Indexed as

EmulsionsOligonucleotide Array Sequence AnalysisCloning, MolecularEscherichia coliFlow CytometryGene ExpressionHigh-Throughput Screening AssaysMicrofluidicsEmulsions

Identifiers

PMID28239692
PMCPMC5428077
OpenAlexW2586871903

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

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.