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Related Concept Videos

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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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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Genome Size and the Evolution of New Genes03:21

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While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.
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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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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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Updated: Jan 28, 2026

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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Base editing the mammalian genome.

Emma M Schatoff1, Maria Paz Zafra2, Lukas E Dow3

  • 1Sandra and Edward Meyer Cancer Center, United States; Weill Cornell/Rockefeller/Sloan Kettering Tri-I MD-PhD Program, New York 10065, United States.

Methods (San Diego, Calif.)
|March 6, 2019
PubMed
Summary
This summary is machine-generated.

Base editing (BE) precisely changes DNA nucleotides in mammals. Recent advancements offer versatile tools for genome modification in research and disease therapy.

Keywords:
APOBECBEBase editingCRISPR

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Area of Science:

  • Genetics
  • Molecular Biology
  • Biotechnology

Background:

  • Base editing (BE) is a genome engineering technology enabling programmable nucleotide conversions.
  • BE systems utilize a modified Cas9 nuclease (Cas9D10A) linked to a DNA-modifying enzyme.
  • BE shows significant potential in basic research and translational medicine for disease allele correction.

Purpose of the Study:

  • To provide an overview of DNA base editing concepts.
  • To discuss recent advancements in optimized base editing systems for mammalian cells.
  • To guide the effective use of base editing tools through technical insights and experimental examples.

Main Methods:

  • Review of base editing technology principles and components.
  • Analysis of recent developments in base editor variants.
  • Presentation of experimental strategies and examples for base editing in cell lines and organoids.

Main Results:

  • BE technology allows precise single nucleotide conversions in the mammalian genome.
  • Numerous new and modified base editor variants have expanded the technology's flexibility.
  • Successful base editing applications demonstrated in cell lines and organoids.

Conclusions:

  • Effective implementation of base editing requires understanding tool advantages and limitations.
  • Optimized base editing systems are advancing for mammalian cell applications.
  • This review provides practical guidance for utilizing base editing in genome modification experiments.