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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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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

CRISPR and crRNAs

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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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Genomics02:02

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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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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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Related Experiment Video

Updated: Feb 1, 2026

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes
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Strategies for Efficient Genome Editing Using CRISPR-Cas9.

Behnom Farboud1, Aaron F Severson2,3,4, Barbara J Meyer5

  • 1Howard Hughes Medical Institute and Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3204.

Genetics
|December 4, 2018
PubMed
Summary
This summary is machine-generated.

CRISPR-Cas9 genome editing efficiency is improved by understanding asymmetric DNA repair. New strategies enhance precise and imprecise editing outcomes in Caenorhabditis elegans.

Keywords:
CRISPR-Cas9Caenorhabditis elegansDNA repairGenome editinghomology-directed repair

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Area of Science:

  • Molecular Biology
  • Genetics
  • Developmental Biology

Background:

  • CRISPR-Cas9 technology revolutionized genome editing across various organisms.
  • Editing efficiency and repair outcomes (precise and imprecise) following double-strand breaks (DSBs) exhibit significant variability.
  • Understanding the mechanisms of DSB repair is crucial for optimizing genome editing strategies.

Purpose of the Study:

  • To investigate the asymmetric nature of DNA repair following DSBs.
  • To develop improved RNA guides and repair templates for enhanced CRISPR-Cas9 editing efficiency.
  • To establish guidelines for predictable and high-frequency precise and imprecise repair outcomes.

Main Methods:

  • Designed RNA guides and repair templates exploiting asymmetric DSB repair.
  • Developed strategies for inserting large exogenous DNA sequences (10 kb) and multiple nucleotide substitutions.
  • Created co-conversion markers for diverse nematode species.
  • Utilized allele-specific Cas9 targets to analyze DSB repair timing, location, frequency, and sex dependence.

Main Results:

  • Demonstrated asymmetric DNA repair favoring one direction for both precise and imprecise repair.
  • Significantly increased the frequency of imprecise insertions/deletions and precise point mutation insertions in Caenorhabditis elegans.
  • Successfully inserted long DNA sequences and multiple nucleotide substitutions distant from DSBs.
  • Identified a striking difference in editing efficiency between maternal and paternal genomes.
  • Observed high-frequency repair events before and after fertilization.

Conclusions:

  • Asymmetric DNA repair provides a basis for enhancing CRISPR-Cas9 editing efficiency.
  • Developed strategies lead to predictable and high-frequency precise and imprecise repair outcomes.
  • The findings offer insights into the timing and mechanisms of DSB repair.
  • Enhanced genome editing tools and strategies are now available for diverse applications in model organisms.