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Phage-mediated Delivery of Targeted sRNA Constructs to Knock Down Gene Expression in E. coli
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Biology
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JoVE Journal Biology
Phage-mediated Delivery of Targeted sRNA Constructs to Knock Down Gene Expression in E. coli

Phage-mediated Delivery of Targeted sRNA Constructs to Knock Down Gene Expression in E. coli

DOI:
10.3791/53618-v

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08:25 min

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March 20, 2016

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, , ,
1U1001,Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes

Chapters

  • 00:05Title
  • 00:40Site Directed Mutagenesis on sRNA Expression Cassette
  • 03:40Production and Harvest of M13-packaged Phagemid Stocks
  • 05:01Silencing by Infection with Packaged Phagemids
  • 06:15Results: Gene Silencing in E. coli Using sRNA Phagemids
  • 07:36Conclusion

Summary

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We describe a method to knock down gene expression in a growing population of E. coli cells using sequence-targeted sRNA expression cassettes delivered by an M13 phagemid vector.

Tags

Phage-mediated DeliveryTargeted SRNA ConstructsGene KnockdownE. ColiGene ExpressionSynthetic BiologyBatch CulturesPlasmidSRNA Expression CassetteSite-directed MutagenesisGuide SequenceForward And Reverse PrimersHomology RegionsTemplate SRNA PhagemidPCR ReactionsHigh Fidelity Polymerase

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