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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

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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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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.
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Related Experiment Video

Updated: Feb 15, 2026

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System
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Efficient CRISPR/Cas9-based genome editing in carrot cells.

Magdalena Klimek-Chodacka1, Tomasz Oleszkiewicz2, Levi G Lowder3

  • 1Institute of Plant Biology and Biotechnology, Faculty of Biotechnology and Horticulture, University of Agriculture in Krakow, AL. 29 Listopada 54, 31-425, Krakow, Poland. m.chodacka@ogr.ur.krakow.pl.

Plant Cell Reports
|January 15, 2018
PubMed
Summary

This study demonstrates successful genome editing in carrots using the CRISPR/Cas9 system. Researchers achieved efficient targeted mutagenesis, paving the way for improved carrot crop development and research.

Keywords:
Daucus carotaFlavanone 3-hydroxylaseGene knockoutGenome editingSite-directed mutagenesis

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Area of Science:

  • Plant Science
  • Molecular Biology
  • Biotechnology

Background:

  • The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas9) system is a powerful genome editing tool.
  • CRISPR/Cas9 has been widely adopted in model organisms but not yet in carrots.
  • Carrots are an important health-promoting crop and a model species for in vitro culture.

Purpose of the Study:

  • To report the first successful and efficient application of the CRISPR/Cas9 system for genome editing in carrots.
  • To target the flavanone-3-hydroxylase (F3H) gene to study anthocyanin biosynthesis.
  • To evaluate different codon-optimized Cas9 genes for efficiency in carrot mutagenesis.

Main Methods:

  • Utilized Agrobacterium-mediated genetic transformation to introduce CRISPR/Cas9 vectors into carrot callus.
  • Employed multiplexing CRISPR/Cas9 vectors with two single-guide RNAs (gRNAs) targeting the F3H gene.
  • Assessed the efficiency of three codon-optimized Cas9 genes, including Arabidopsis codon-optimized AteCas9.

Main Results:

  • Achieved up to 90% efficiency in generating F3H mutants using the AteCas9 gene.
  • Observed discoloration of calli upon F3H gene knockout, confirming its role in anthocyanin biosynthesis.
  • Generated small Indels and long chromosome fragment deletions (116-119 nt) via simultaneous cleavage.

Conclusions:

  • Demonstrated successful site-directed mutagenesis in carrot using the CRISPR/Cas9 system.
  • Validated the utility of a model callus culture for assessing genome editing systems.
  • Highlighted the potential of genome editing for advancing carrot research and crop improvement.