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Related Concept Videos

Genomic DNA in Prokaryotes00:46

Genomic DNA in Prokaryotes

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The genome of most prokaryotic organisms consists of double-stranded DNA organized into one circular chromosome in a region of cytoplasm called the nucleoid. The chromosome is tightly wound, or supercoiled, for efficient storage. Prokaryotes also contain other circular pieces of DNA called plasmids. These plasmids are smaller than the chromosome and often carry genes that confer adaptive functions, such as antibiotic resistance.
Genomic Diversity in Bacteria
Although bacterial genomes are much...
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Prokaryotic Cells01:51

Prokaryotic Cells

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Prokaryotes are small unicellular organisms that include the domains—Archaea and Bacteria. Bacteria include many common organisms, such as Salmonella and E. coli, while the Archaea include extremophiles that live in harsh environments, such as volcanic springs.
Like eukaryotic cells, all prokaryotic cells are surrounded by a plasma membrane, have genetic material in the form of single, circular DNA, a cytoplasm that fills the interior of the cell, and ribosomes that synthesize proteins....
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Bacterial Transformation01:33

Bacterial Transformation

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In 1928, bacteriologist Frederick Griffith worked on a vaccine for pneumonia, which is caused by Streptococcus pneumoniae bacteria. Griffith studied two pneumonia strains in mice: one pathogenic and one non-pathogenic. Only the pathogenic strain killed host mice.
Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...
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Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

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The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...
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Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

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Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
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Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
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Targeted, Genome-scale Overexpression in Proteobacteria.

Amy B Banta1,2, Kevin S Myers1,2, Ryan D Ward2,3

  • 1Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison, Madison, WI, USA.

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Summary

Researchers developed CRISPRtOE, a new method for genome-scale gene overexpression in bacteria. This tool enables enhanced gene expression for discovering antibiotic targets and optimizing industrial strains.

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A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae
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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Genome-scale gene perturbation screens using CRISPR interference (CRISPRi) and activation (CRISPRa) are vital in eukaryotic research.
  • While CRISPRi knockdowns are common in bacteria, efficient genome-scale gene overexpression methods are limited.

Conclusions:

  • CRISPRtOE is a powerful new tool for bacterial genetics.
  • Enables advancements in understanding antibiotic mechanisms, optimizing industrial microbial strains, and discovering novel gene functions.