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Related Concept Videos

CRISPR01:59

CRISPR

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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/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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What is Genetic Engineering?00:49

What is Genetic Engineering?

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Overview
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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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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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Genomic Imprinting and Inheritance02:30

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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.
The expression of some genes depends on which parent passed the gene to the offspring, through a phenomenon known as...
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Genome Engineering of Primary Human B Cells Using CRISPR/Cas9
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Genome Engineering of Primary Human B Cells Using CRISPR/Cas9

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Efficient mouse genome engineering by CRISPR-EZ technology.

Andrew J Modzelewski1, Sean Chen1, Brandon J Willis2

  • 1Division of Cellular and Developmental Biology, Department of Molecular & Cell Biology, University of California, Berkeley, Berkeley, California, USA.

Nature Protocols
|May 12, 2018
PubMed
Summary
This summary is machine-generated.

CRISPR-EZ electroporation offers a faster, simpler, and more efficient method for mouse genome editing compared to microinjection. This new technique enables rapid generation of genetically modified mice, streamlining research workflows.

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Mouse Genome Engineering Using Designer Nucleases
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Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes
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Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 technology revolutionized genome editing but microinjection is a bottleneck.
  • Efficient and high-throughput mouse genome engineering is crucial for research.

Purpose of the Study:

  • To develop and optimize a novel electroporation-based method for CRISPR/Cas9 genome editing in mouse zygotes.
  • To compare the efficiency, simplicity, cost, and throughput of the new method against traditional microinjection.

Main Methods:

  • Developed CRISPR ribonucleoprotein (RNP) electroporation of zygotes (CRISPR-EZ).
  • Optimized sgRNA synthesis, embryo collection, RNP electroporation, and genotyping.
  • Conducted side-by-side comparisons with microinjection in C57BL/6J and C57BL/6N mouse strains within the KnockOut Mouse Project (KOMP) pipeline.

Main Results:

  • CRISPR-EZ demonstrated 100% delivery of Cas9/sgRNA RNPs in mouse zygotes.
  • Achieved high efficiency for various genome modifications including indels, exon deletions, point mutations, and small insertions.
  • CRISPR-EZ consistently outperformed microinjection in efficiency, simplicity, cost, and throughput.

Conclusions:

  • CRISPR-EZ is a superior alternative to microinjection for mouse genome engineering.
  • This technology simplifies and accelerates the generation of genetically modified mice, making it accessible to researchers with basic skills.
  • CRISPR-EZ holds potential for application in other mammalian species.