Multiplexed gene synthesis in emulsions for exploring protein functional landscapes
Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S.
American Association for the Advancement of Science (AAAS)
Published 2018
American Association for the Advancement of Science (AAAS)
Published 2018
| Publication Date: |
2018-01-19
|
|---|---|
| Publisher: |
American Association for the Advancement of Science (AAAS)
|
| Print ISSN: |
0036-8075
|
| Electronic ISSN: |
1095-9203
|
| Topics: |
Biology
Chemistry and Pharmacology
Geosciences
Computer Science
Medicine
Natural Sciences in General
Physics
|
| Keywords: |
Molecular Biology
|
| Published by: |
| _version_ | 1836398756577148928 |
|---|---|
| autor | Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. |
| beschreibung | Improving our ability to construct and functionally characterize DNA sequences would broadly accelerate progress in biology. Here, we introduce DropSynth, a scalable, low-cost method to build thousands of defined gene-length constructs in a pooled (multiplexed) manner. DropSynth uses a library of barcoded beads that pull down the oligonucleotides necessary for a gene’s assembly, which are then processed and assembled in water-in-oil emulsions. We used DropSynth to successfully build more than 7000 synthetic genes that encode phylogenetically diverse homologs of two essential genes in Escherichia coli . We tested the ability of phosphopantetheine adenylyltransferase homologs to complement a knockout E. coli strain in multiplex, revealing core functional motifs and reasons underlying homolog incompatibility. DropSynth coupled with multiplexed functional assays allows us to rationally explore sequence-function relationships at an unprecedented scale. |
| citation_standardnr | 6142338 |
| datenlieferant | ipn_articles |
| feed_id | 25 |
| feed_publisher | American Association for the Advancement of Science (AAAS) |
| feed_publisher_url | http://www.aaas.org/ |
| insertion_date | 2018-01-19 |
| journaleissn | 1095-9203 |
| journalissn | 0036-8075 |
| publikationsjahr_anzeige | 2018 |
| publikationsjahr_facette | 2018 |
| publikationsjahr_intervall | 7984:2015-2019 |
| publikationsjahr_sort | 2018 |
| publisher | American Association for the Advancement of Science (AAAS) |
| quelle | Science |
| relation | http://science.sciencemag.org/cgi/content/short/359/6373/343?rss=1 |
| schlagwort | Molecular Biology |
| search_space | articles |
| shingle_author_1 | Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. |
| shingle_author_2 | Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. |
| shingle_author_3 | Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. |
| shingle_author_4 | Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. |
| shingle_catch_all_1 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes Molecular Biology Improving our ability to construct and functionally characterize DNA sequences would broadly accelerate progress in biology. Here, we introduce DropSynth, a scalable, low-cost method to build thousands of defined gene-length constructs in a pooled (multiplexed) manner. DropSynth uses a library of barcoded beads that pull down the oligonucleotides necessary for a gene’s assembly, which are then processed and assembled in water-in-oil emulsions. We used DropSynth to successfully build more than 7000 synthetic genes that encode phylogenetically diverse homologs of two essential genes in Escherichia coli . We tested the ability of phosphopantetheine adenylyltransferase homologs to complement a knockout E. coli strain in multiplex, revealing core functional motifs and reasons underlying homolog incompatibility. DropSynth coupled with multiplexed functional assays allows us to rationally explore sequence-function relationships at an unprecedented scale. Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. American Association for the Advancement of Science (AAAS) 0036-8075 00368075 1095-9203 10959203 |
| shingle_catch_all_2 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes Molecular Biology Improving our ability to construct and functionally characterize DNA sequences would broadly accelerate progress in biology. Here, we introduce DropSynth, a scalable, low-cost method to build thousands of defined gene-length constructs in a pooled (multiplexed) manner. DropSynth uses a library of barcoded beads that pull down the oligonucleotides necessary for a gene’s assembly, which are then processed and assembled in water-in-oil emulsions. We used DropSynth to successfully build more than 7000 synthetic genes that encode phylogenetically diverse homologs of two essential genes in Escherichia coli . We tested the ability of phosphopantetheine adenylyltransferase homologs to complement a knockout E. coli strain in multiplex, revealing core functional motifs and reasons underlying homolog incompatibility. DropSynth coupled with multiplexed functional assays allows us to rationally explore sequence-function relationships at an unprecedented scale. Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. American Association for the Advancement of Science (AAAS) 0036-8075 00368075 1095-9203 10959203 |
| shingle_catch_all_3 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes Molecular Biology Improving our ability to construct and functionally characterize DNA sequences would broadly accelerate progress in biology. Here, we introduce DropSynth, a scalable, low-cost method to build thousands of defined gene-length constructs in a pooled (multiplexed) manner. DropSynth uses a library of barcoded beads that pull down the oligonucleotides necessary for a gene’s assembly, which are then processed and assembled in water-in-oil emulsions. We used DropSynth to successfully build more than 7000 synthetic genes that encode phylogenetically diverse homologs of two essential genes in Escherichia coli . We tested the ability of phosphopantetheine adenylyltransferase homologs to complement a knockout E. coli strain in multiplex, revealing core functional motifs and reasons underlying homolog incompatibility. DropSynth coupled with multiplexed functional assays allows us to rationally explore sequence-function relationships at an unprecedented scale. Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. American Association for the Advancement of Science (AAAS) 0036-8075 00368075 1095-9203 10959203 |
| shingle_catch_all_4 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes Molecular Biology Improving our ability to construct and functionally characterize DNA sequences would broadly accelerate progress in biology. Here, we introduce DropSynth, a scalable, low-cost method to build thousands of defined gene-length constructs in a pooled (multiplexed) manner. DropSynth uses a library of barcoded beads that pull down the oligonucleotides necessary for a gene’s assembly, which are then processed and assembled in water-in-oil emulsions. We used DropSynth to successfully build more than 7000 synthetic genes that encode phylogenetically diverse homologs of two essential genes in Escherichia coli . We tested the ability of phosphopantetheine adenylyltransferase homologs to complement a knockout E. coli strain in multiplex, revealing core functional motifs and reasons underlying homolog incompatibility. DropSynth coupled with multiplexed functional assays allows us to rationally explore sequence-function relationships at an unprecedented scale. Plesa, C., Sidore, A. M., Lubock, N. B., Zhang, D., Kosuri, S. American Association for the Advancement of Science (AAAS) 0036-8075 00368075 1095-9203 10959203 |
| shingle_title_1 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes |
| shingle_title_2 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes |
| shingle_title_3 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes |
| shingle_title_4 | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes |
| timestamp | 2025-06-30T23:32:08.169Z |
| titel | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes |
| titel_suche | Multiplexed gene synthesis in emulsions for exploring protein functional landscapes |
| topic | W V TE-TZ SQ-SU WW-YZ TA-TD U |
| uid | ipn_articles_6142338 |