Droplet microfluidics for synthetic biology

Gach, Philip C.; Iwai, Kosuke; Kim, Peter W.; Hillson, Nathan J.; Singh, Anup K.

doi:10.1039/c7lc00576h

Cited by 102 publications

(56 citation statements)

References 125 publications

(108 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This especially concerns applications such as droplet‐based cell sorting, drug delivery, DNA sequencing, or material synthesis . In particular, the field of bottom‐up synthetic biology will benefit from the dynamic control over the organization of intracellular components . These applications require mechanisms to turn interactions with the droplet periphery on and off upon demand.…”

Section: Resultsmentioning

confidence: 99%

Programmable Functionalization of Surfactant‐Stabilized Microfluidic Droplets via DNA‐Tags

Jahnke

Weiss

Frey

et al. 2019

Adv Funct Materials

View full text Add to dashboard Cite

Droplet-based microfluidics has emerged as a powerful tool in synthetic biology. For many applications, chemical functionalization of the droplets is a key process. Therefore, a straightforward and broadly applicable approach is developed to functionalize the inner periphery of microfluidic droplets with diverse reactive groups and components. Instead of covalent modification of the droplet-stabilizing surfactants, this method relies on cholesteroltagged DNA that self-assembles at the droplet periphery. The cholesteroltagged DNA serves as an attachment handle for the recruitment of complementary DNA. The complementary DNA can carry diverse functional groups. We exemplify our method by demonstrating the attachment of amine groups, DNA nanostructures, microspheres, a minimal actin cortex, and leukemia cells to the droplet periphery. It is further shown that the DNAmediated attachment to the droplet periphery is temperature-responsive and reversible. It is envisioned that droplet functionalization via DNA handles will help to tailor droplet interfaces for diverse applications-featuring programmable assembly, unique addressability, and stimuli-responsiveness.The ORCID identification number(s) for the author(s) of this article can be found under https://doi.org/10.1002/adfm.201808647. not universally applicable-it depends on the success of the chemical synthesis and can interfere with the stability and the physical properties of the droplets. Moreover, the binding of the functional group to the droplet periphery is irreversible.DNA nanotechnology, [12] on the other hand can attain the programmable assembly of arbitrary nanoscale architectures like DNA-based lattices, [13] nanopores, [14][15][16] or lid-containing boxes. [17,18] DNA has also been used as a scaffold or linker to assemble secondary components including proteins, [19] gold nanoparticles [20] and liposomes. [21] In addition, networks of emulsion droplets [22,23] or colloid-coated droplets [24] have been created using DNA linkers. Yet in all cases, the linkage was based on biotinylated DNA, which requires additional efforts to graft streptavidin onto the droplet surface. Furthermore, it has never been demonstrated that it is possible to functionalize the interior of block-copolymer surfactantstabilized droplets with DNA.Here, we present a broadly applicable method for functionalizing microfluidic droplets utilizing the hydrophobic interaction of cholesterol-tagged DNA with the droplet-stabilizing surfactant. Notably, the interaction of cholesterol with perfluorinated chains has never been described or exploited before. We show that DNA handles can serve as reversible anchoring points for various components including reactive groups, DNA nanostructures, beads, proteins or even cells. The use of off-the-shelf available DNA holds considerable advantages compared to standard methods for droplet functionalization, including: the broad scope of options for site-directed chemical functionalization, the addressability and programmability due to specific base pairin...

show abstract

Section: Resultsmentioning

confidence: 99%

Programmable Functionalization of Surfactant‐Stabilized Microfluidic Droplets via DNA‐Tags

Jahnke

Weiss

Frey

et al. 2019

Adv Funct Materials

View full text Add to dashboard Cite

show abstract

“…However, recently a high-throughput, droplet-based microfluidic method was developed to generate defined-size vesicles termed droplet-stabilized GUVs. [119][120][121][122] In general, an increase in complexity may be necessary to eventually reach the ultimate aim to design a minimal cell. [118] This highlights the potential of microfluidic techniques, which enables not only for creation of a complex biomimetic microcompartment, but also for the assembly of biological modules to a functional system-a task that could further be complemented by advances in 3D-printing for future attempts of minimal cell assembly.…”

Section: Toward Minimal Cell Design: Compartmentalization and Increasmentioning

confidence: 99%

“…[118] This highlights the potential of microfluidic techniques, which enables not only for creation of a complex biomimetic microcompartment, but also for the assembly of biological modules to a functional system-a task that could further be complemented by advances in 3D-printing for future attempts of minimal cell assembly. [119][120][121][122] In general, an increase in complexity may be necessary to eventually reach the ultimate aim to design a minimal cell. The challenge here is to keep a balance between the level of complexity needed for certain processes and holding the upper hand over controlling these processes.…”

Section: Toward Minimal Cell Design: Compartmentalization and Increasmentioning

confidence: 99%

Cytoskeletal and Actin‐Based Polymerization Motors and Their Role in Minimal Cell Design

2019

View full text Add to dashboard Cite

Life implies motion. In cells, protein‐based active molecular machines drive cell locomotion and intracellular transport, control cell shape, segregate genetic material, and split a cell in two parts. Key players among molecular machines driving these various cell functions are the cytoskeleton and motor proteins that convert chemical bound energy into mechanical work. Findings over the last decades in the field of in vitro reconstitutions of cytoskeletal and motor proteins have elucidated mechanistic details of these active protein systems. For example, a complex spatial and temporal interplay between the cytoskeleton and motor proteins is responsible for the translation of chemically bound energy into (directed) movement and force generation, which eventually governs the emergence of complex cellular functions. Understanding these mechanisms and the design principles of the cytoskeleton and motor proteins builds the basis for mimicking fundamental life processes. Here, a brief overview of actin, prokaryotic actin analogs, and motor proteins and their potential role in the design of a minimal cell from the bottom‐up is provided.

show abstract

“…There are numerous reviews on droplet-based microfluidic systems focusing on droplet generation [9][10][11], droplet manipulation [12][13][14] and applications such as single cell analysis [15,16], biochemical detection [17] and synthetic biology [18]. Specifically, this review aims to summarize the label-free sensing systems demonstrated using microdroplets.…”

Section: Introductionmentioning

confidence: 99%

Label-Free Sensing in Microdroplet-Based Microfluidic Systems

2018

View full text Add to dashboard Cite

Droplet microfluidic systems have evolved as fluidic platforms that use much less sample volume and provide high throughput for biochemical analysis compared to conventional microfluidic devices. The variety of droplet fluidic applications triggered several detection techniques to be applied for analysis of droplets. In this review, we focus on label-free droplet detection techniques that were adapted to various droplet microfluidic platforms. We provide a classification of most commonly used droplet platform technologies. Then we discuss the examples of various label-free droplet detection schemes implemented for these platforms. While providing the research landscape for label-free droplet detection methods, we aim to highlight the strengths and shortcomings of each droplet platform so that a more targeted approach can be taken by researchers when selecting a droplet platform and a detection scheme for any given application.

show abstract

Droplet microfluidics for synthetic biology

Abstract: Review of current droplet microfluidics systems as they apply to the field of synthetic biology and genetic engineering.

Cited by 102 publications

References 125 publications

Programmable Functionalization of Surfactant‐Stabilized Microfluidic Droplets via DNA‐Tags

Programmable Functionalization of Surfactant‐Stabilized Microfluidic Droplets via DNA‐Tags

Cytoskeletal and Actin‐Based Polymerization Motors and Their Role in Minimal Cell Design

Label-Free Sensing in Microdroplet-Based Microfluidic Systems

Contact Info

Product

Resources

About