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Related Concept Videos

Induced Pluripotent Stem Cells01:13

Induced Pluripotent Stem Cells

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Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore...
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Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
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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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Embryonic Stem Cells00:58

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Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.
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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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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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Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing
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Efficient scarless genome editing in human pluripotent stem cells.

Kazuya Ikeda1, Nobuko Uchida2, Toshinobu Nishimura3

  • 1Department of Pediatrics, Stanford University, Stanford, CA, USA.

Nature Methods
|December 4, 2018
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Achieve scarless genome editing in human pluripotent stem cells (hPSCs) using a novel CRISPR-Cas9 method. This technique combines homologous recombination with selection to enable precise genetic modifications for research and therapies.

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Area of Science:

  • Biotechnology
  • Stem Cell Biology
  • Gene Editing

Background:

  • Scarless genome editing in human pluripotent stem cells (hPSCs) is crucial for therapeutic applications.
  • Current methods may leave genetic scars, limiting clinical translation.

Purpose of the Study:

  • To develop a versatile and efficient method for scarless genome editing in hPSCs.
  • To enable precise genetic modifications for research and clinical use.

Main Methods:

  • Utilized CRISPR-Cas9 technology for targeted gene modification.
  • Employed homologous recombination for precise DNA insertion or correction.
  • Implemented a positive-negative selection strategy to isolate accurately edited clones.

Main Results:

  • Successfully generated scarless genetic modifications in hPSCs.
  • Demonstrated the efficiency and versatility of the combined approach.
  • Established a robust method for producing genetically precise hPSC lines.

Conclusions:

  • The developed method offers a significant advancement for scarless genome editing in hPSCs.
  • This technique facilitates the clinical translation of hPSC-derived therapies.
  • Enables precise genetic engineering for diverse research applications.