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Related Concept Videos

Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

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As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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Overview of Transposition and Recombination02:13

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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Translation01:31

Translation

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Lesson: Translation
Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA), and other proteins produce a chain of amino acids—the polypeptide—as the end product of translation.
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Translation01:31

Translation

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Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA), and other proteins produce a chain of amino acids—the polypeptide—as the end product of translation.
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae
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TALEN-Induced Translocations in Human Cells.

Marion Piganeau1,2,3, Benjamin Renouf1,2,3, Hind Ghezraoui1,2,3

  • 1Museum National d'Histoire Naturelle, 43 rue Cuvier, 75005, Paris, France.

Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2015
PubMed
Summary

This study details a method for inducing chromosomal translocations in human cells using transcription activator-like effector nucleases (TALENs) to investigate cancer and genome instability. The described techniques allow for detection, frequency calculation, and breakpoint analysis of these translocations.

Keywords:
96-Well plate PCR screenBreakpoint junction analysisFusion geneT7-endonuclease assayTALEN induced translocation

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Area of Science:

  • Genetics
  • Molecular Biology
  • Cancer Research

Background:

  • Chromosomal translocations are key drivers of tumorigenesis and indicators of genome instability.
  • Understanding translocation mechanisms is crucial for cancer research and developing therapeutic strategies.

Purpose of the Study:

  • To provide a detailed methodology for inducing and analyzing chromosomal translocations in human cells.
  • To establish reliable methods for detecting and quantifying translocation formation and characterizing breakpoint junctions.

Main Methods:

  • Utilizing transcription activator-like effector nucleases (TALENs) to induce targeted chromosomal translocations.
  • Employing Polymerase Chain Reaction (PCR) for translocation detection and a 96-well PCR screen for frequency calculation.
  • Analyzing breakpoint junctions and detecting fusion genes via Fluorescence In Situ Hybridization (FISH) or Western Blot analysis.

Main Results:

  • Successful induction of chromosomal translocations in human cells using the TALENs system.
  • Established protocols for accurate detection, frequency determination, and detailed analysis of translocation breakpoints.
  • Demonstrated feasibility of detecting cancer-relevant fusion genes.

Conclusions:

  • The TALENs-based method offers a robust approach for studying chromosomal translocations and their role in cancer.
  • The described techniques facilitate comprehensive analysis of translocation formation, frequency, and molecular characteristics.
  • This methodology aids in advancing research on genome instability and tumorigenesis.