WO2016048994A3 - Systems and methods of cell-free protein synthesis in droplets and other compartments - Google Patents
Systems and methods of cell-free protein synthesis in droplets and other compartments Download PDFInfo
- Publication number
- WO2016048994A3 WO2016048994A3 PCT/US2015/051401 US2015051401W WO2016048994A3 WO 2016048994 A3 WO2016048994 A3 WO 2016048994A3 US 2015051401 W US2015051401 W US 2015051401W WO 2016048994 A3 WO2016048994 A3 WO 2016048994A3
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- protein
- droplets
- cell
- free
- protein synthesis
- Prior art date
Links
- 238000001243 protein synthesis Methods 0.000 title abstract 4
- 230000014616 translation Effects 0.000 title abstract 4
- 238000000034 method Methods 0.000 title 1
- 102000004169 proteins and genes Human genes 0.000 abstract 6
- 108090000623 proteins and genes Proteins 0.000 abstract 6
- 230000002068 genetic effect Effects 0.000 abstract 3
- 102000039446 nucleic acids Human genes 0.000 abstract 2
- 108020004707 nucleic acids Proteins 0.000 abstract 2
- 150000007523 nucleic acids Chemical class 0.000 abstract 2
- 238000010396 two-hybrid screening Methods 0.000 abstract 2
- 238000003556 assay Methods 0.000 abstract 1
- 239000011230 binding agent Substances 0.000 abstract 1
- 210000004671 cell-free system Anatomy 0.000 abstract 1
- 230000000694 effects Effects 0.000 abstract 1
- 102000037865 fusion proteins Human genes 0.000 abstract 1
- 108020001507 fusion proteins Proteins 0.000 abstract 1
- 238000013537 high throughput screening Methods 0.000 abstract 1
- 238000000338 in vitro Methods 0.000 abstract 1
- 230000004850 protein–protein interaction Effects 0.000 abstract 1
- 238000012216 screening Methods 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/80—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention generally relates to cell-free protein synthesis in microfluidic droplets. Droplets may be used to encapsulate genetic information (DNA/RNA), and through cell-free protein synthesis, provide a linkage of the genetic information with the functional information, e.g., the activity of the expressed protein, and used to convert the functional information into a detectable signal, e.g., to allow sorting of the droplets and retrieve genetic information associated with the drops. In one set of embodiments, microfluidic droplets containing a cell-free protein synthesis system designed to detect protein-protein interaction (e.g., in vitro two-hybrid systems or IVT2H) can be used for high-throughput screening, e.g., of protein binders. This drop-based two-hybrid system may include two (or more) fusion proteins that can bind to each other such that their binding produces a complex that is able to produce a nucleic acid. The nucleic acid may be expressed to produce a protein. In certain embodiments, the protein may be produced within a cell-free system. The protein may be fluorescent or otherwise determinable, such that determination of the protein may be used to allow assays to occur within the droplets, to allow sorting of the droplets to occur, or the like, e.g., as discussed below, for instance, for screening or other applications.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462054263P | 2014-09-23 | 2014-09-23 | |
| US62/054,263 | 2014-09-23 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO2016048994A2 WO2016048994A2 (en) | 2016-03-31 |
| WO2016048994A3 true WO2016048994A3 (en) | 2016-05-26 |
Family
ID=55582238
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2015/051401 WO2016048994A2 (en) | 2014-09-23 | 2015-09-22 | Systems and methods of cell-free protein synthesis in droplets and other compartments |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2016048994A2 (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018067792A1 (en) * | 2016-10-07 | 2018-04-12 | President And Fellows Of Harvard College | Sequencing of bacteria or other species |
| EP3828283A1 (en) * | 2019-11-28 | 2021-06-02 | Diagenode S.A. | An improved sequencing method and kit |
| US11788137B2 (en) | 2019-09-30 | 2023-10-17 | Diagenode S.A. | Diagnostic and/or sequencing method and kit |
| GB202311993D0 (en) * | 2023-08-04 | 2023-09-20 | Cambridge Entpr Ltd | Method and system |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030008398A1 (en) * | 1996-12-11 | 2003-01-09 | Rolf Mueller | Self-enhancing, pharmacologically controllable expression systems |
| US20090304699A1 (en) * | 2006-05-24 | 2009-12-10 | Biogen Idec Ma Inc. | Methods of treating fibrosis |
| WO2012139134A2 (en) * | 2011-04-07 | 2012-10-11 | Coferon, Inc. | Methods of modulating oncogenic fusion proteins |
| US20130171328A1 (en) * | 2010-06-02 | 2013-07-04 | Ganesh M. Kishore | Production of steviol glycosides in microorganisms |
-
2015
- 2015-09-22 WO PCT/US2015/051401 patent/WO2016048994A2/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030008398A1 (en) * | 1996-12-11 | 2003-01-09 | Rolf Mueller | Self-enhancing, pharmacologically controllable expression systems |
| US20090304699A1 (en) * | 2006-05-24 | 2009-12-10 | Biogen Idec Ma Inc. | Methods of treating fibrosis |
| US20130171328A1 (en) * | 2010-06-02 | 2013-07-04 | Ganesh M. Kishore | Production of steviol glycosides in microorganisms |
| WO2012139134A2 (en) * | 2011-04-07 | 2012-10-11 | Coferon, Inc. | Methods of modulating oncogenic fusion proteins |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016048994A2 (en) | 2016-03-31 |
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