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Related Concept Videos

Genomics02:02

Genomics

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Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...
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CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Genome Size and the Evolution of New Genes03:21

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While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.
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Diploid organisms inherit genetic material through chromosomes from both parents. Copies of the same gene are known as alleles. In most cases, both alleles are simultaneously expressed and allow various cellular processes to function optimally. If one of the alleles is missing or mutated, the expression of the other allele can compensate; however, this is not true for all genes.
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Comparing Mitochondrial, Chloroplast, and Prokaryotic Genomes02:16

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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
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Pooled CRISPR interference screening enables genome-scale functional genomics study in bacteria with superior

Tianmin Wang1, Changge Guan1, Jiahui Guo1

  • 1MOE Key Laboratory for Industrial Biocatalysis, Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China.

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|June 28, 2018
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We developed a CRISPR interference (CRISPRi) pooled screening method for bacteria to link genes with cellular functions. This high-throughput tool precisely maps microbial genetic networks and outperforms existing methods.

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Area of Science:

  • Microbial Genomics
  • Synthetic Biology
  • Functional Genomics

Background:

  • Exploiting microbial genome resources requires high-throughput methods to link genes with phenotypes.
  • Existing methods like transposon sequencing have limitations in microbial functional genomics.

Purpose of the Study:

  • To develop a novel CRISPR interference (CRISPRi) pooled screening method for bacteria.
  • To enable genome-level association of genes with cellular phenotypes.
  • To establish a powerful tool for mapping prokaryotic genetic networks.

Main Methods:

  • Designed a genome-scale guide RNA library (~60,000 members) for E. coli based on tiling screening.
  • Applied CRISPR interference (CRISPRi) pooled screening to investigate gene repression consequences.
  • Developed rules for guide RNA library design tailored to prokaryotic genomes.

Main Results:

  • CRISPRi pooled screening effectively links genes to phenotypes at the genome level.
  • The method outperforms transposon sequencing for microbial functional genomics, especially for short genes.
  • A comprehensive tRNA-fitness map was constructed, demonstrating utility for non-coding RNAs (ncRNAs).

Conclusions:

  • CRISPRi pooled screening is a precise and high-throughput tool for microbial functional genomics.
  • This method advances the study of prokaryotic genetic networks and ncRNAs.
  • The developed platform facilitates full exploitation of microbial genome resources.