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RNA Editing02:23

RNA Editing

9.9K
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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Plasmids01:28

Plasmids

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Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...
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Cell Specific Gene Expression01:58

Cell Specific Gene Expression

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Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
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Cell Specific Gene Expression01:58

Cell Specific Gene Expression

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Gene Families01:57

Gene Families

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Gene families consist of groups of genes proposed to have originated from a common ancestor. Typically these arise through events in which a gene or genes are mistakenly duplicated during cell division. Unlike their parent genes (which are subject to selection pressure to maintain function), these gene copies do not need to preserve their sequences and may evolve at a relatively faster rate.
Occasionally these regions can be adapted to take on new roles within the organism, becoming novel genes...
10.0K
Combinatorial Gene Control02:33

Combinatorial Gene Control

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Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
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Related Experiment Video

Updated: Feb 14, 2026

Gene Editing of Primary Rhesus Macaque B Cells
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Gene Editing of Primary Rhesus Macaque B Cells

Published on: February 10, 2023

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Highly Efficient and Versatile Plasmid-Based Gene Editing in Primary T Cells.

Mara Kornete1, Romina Marone1, Lukas T Jeker2

  • 1Department of Biomedicine, Basel University Hospital and University of Basel, CH-4031 Basel, Switzerland; and Transplantation Immunology and Nephrology, Basel University Hospital, CH-4031 Basel, Switzerland.

Journal of Immunology (Baltimore, Md. : 1950)
|February 16, 2018
PubMed
Summary

This study introduces an efficient CRISPR gene editing system for primary T cells. The method enables precise genetic modifications, including gene ablation and mutation correction, advancing cellular therapies.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • Adoptive cell transfer is a key research tool and therapeutic strategy for diseases like infections and blood cancers.
  • Genetic manipulation of primary immune cells offers novel research avenues and potential enhancements for cellular therapies.

Purpose of the Study:

  • To develop a highly efficient genome engineering system for primary murine T cells using a CRISPR-Cas9 approach.
  • To demonstrate the system's capability for gene ablation, precise single nucleotide polymorphism introduction, and disease-causing mutation correction.

Main Methods:

  • Utilized a plasmid-based RNA-guided CRISPR system for genome engineering in primary murine T cells.
  • Employed gene editing-mediated allele switching to quantify homology-directed repair and optimize experimental parameters.
  • Applied the system to correct mutations in the Foxp3 gene associated with scurfy syndrome and a clinical mutation.

Main Results:

  • Achieved high efficiency gene ablation (up to 90%) and targeted single nucleotide polymorphism introduction (up to 25%) in primary T cells.
  • Successfully mapped a B cell epitope and demonstrated enrichment of edited cells using allele switching.
  • Restored Foxp3 expression in primary T cells by correcting disease-causing mutations, including a 2-bp insertion and the Foxp3K276X mutation.

Conclusions:

  • The developed CRISPR system provides a straightforward and efficient method for genome engineering in primary T cells.
  • This technology has significant implications for basic research and the development of enhanced cellular therapeutics.
  • The ability to correct disease-causing mutations offers potential for treating genetic disorders affecting T cell function.