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  Imaging-based screening platform assists protein engineering

Fabritius, A., Ng, D., Kist, A. M., Erdogan, M., Portugues, R., & Griesbeck, O. (2018). Imaging-based screening platform assists protein engineering. Cell Chemical Biology, 25(12), 1554-1561.e8. doi:10.1016/j.chembiol.2018.08.008.

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Genre: Journal Article

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 Creators:
Fabritius, Arne1, Author           
Ng, David1, Author           
Kist, Andreas Michael2, Author           
Erdogan, Mutlu1, Author           
Portugues, Ruben2, Author           
Griesbeck, Oliver1, Author           
Affiliations:
1Research Group: Tools for Bio-Imaging / Griesbeck, MPI of Neurobiology, Max Planck Society, ou_1113560              
2Max Planck Research Group: Sensorimotor Control / Portugues, MPI of Neurobiology, Max Planck Society, ou_2054291              

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Free keywords: CYAN FLUORESCENT PROTEIN; DIRECTED EVOLUTION; BRIGHT; MICROSCOPY; ZEBRAFISH; NEURONSBiochemistry & Molecular Biology;
 Abstract: Protein engineering involves generating and screening large numbers of variants for desired properties. While modern DNA technology has made it easy to create protein diversity on the DNA level, the selection and validation of candidate proteins from large libraries remains a challenge. We built a screening platform that integrates high-quality fluorescence-based image analysis and robotic picking of bacterial colonies. It allows tracking each individual colony in a large population and collecting quantitative information on library composition during the protein evolution process. We demonstrate the power of the screening platform by optimizing a dim far-red-emitting fluorescent protein whose brightness increased several fold using iterative cycles of mutagenesis and platform-based screening. The resulting protein variant mCarmine is useful for imaging cells and structures within live tissue as well as for molecular tagging. Overall, the platform presented provides powerful, flexible, and low-cost instrumentation to accelerate many fluorescence-based protein optimization projects.

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Language(s): eng - English
 Dates: 2018
 Publication Status: Published in print
 Pages: 16
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: Cell Chemical Biology
Source Genre: Journal
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Publ. Info: Cell Press
Pages: - Volume / Issue: 25 (12) Sequence Number: - Start / End Page: 1554 - 1561.e8 Identifier: ISSN: 2451-9456
CoNE: https://pure.mpg.de/cone/journals/resource/2451-9456