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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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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Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
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CRISPR01:59

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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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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RNA Editing02:23

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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A Multiplexed CRISPR/Cas9 Editing System Based on the Endogenous tRNA Processing.

Kabin Xie1, Yinong Yang2

  • 1National Key Laboratory for Crop Genetic Improvement and Plant Gene Research Center (Wuhan), Huazhong Agricultural University, Wuhan, China.

Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method for multiplex genome editing using CRISPR-Cas9 technology. This efficient polycistronic tRNA-gRNA (PTG) system enables the simultaneous expression of multiple guide RNAs for precise genetic modifications.

Keywords:
CRISPR-Cas9Genome editingMultiplexgRNAtRNA

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas9 is a widely used genome editing tool known for its efficiency.
  • Multiplex genome editing, requiring multiple guide RNAs (gRNAs), is crucial for complex genetic studies.
  • Simultaneous gRNA expression is a key challenge in multiplex CRISPR applications.

Purpose of the Study:

  • To present a general and efficient method for producing multiple gRNAs from a single transcript.
  • To enable robust multiplex genome editing across diverse species.
  • To provide a detailed protocol for constructing polycistronic tRNA-gRNA (PTG) genes.

Main Methods:

  • Utilized the endogenous tRNA-processing system for gRNA expression.
  • Designed and constructed polycistronic tRNA-gRNA (PTG) genes.
  • Validated the PTG method in various plant, animal, and microbial species.

Main Results:

  • Demonstrated efficient production of multiple gRNAs from a single gene transcript.
  • Achieved highly efficient multiplex genome editing using the PTG system.
  • Confirmed the versatility of the PTG method across different biological systems.

Conclusions:

  • The PTG method offers an efficient strategy for multiplex genome editing.
  • This approach simplifies the simultaneous expression of multiple gRNAs.
  • The PTG system is a valuable tool for advancing genome editing applications in diverse organisms.