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Related Concept Videos

Genetic Screens02:46

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
In vitro Mutagenesis01:16

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
Spontaneous and Induced Mutations01:30

Spontaneous and Induced Mutations

Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
Mismatch Repair01:20

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
Mutations01:35

Mutations

Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations can be characterized in a number of different ways: whether and how they alter the amino acid sequence of the protein, whether they occur over a small or large area of DNA, and whether they occur in somatic cells or germline cells.
Chromosomal Alterations Are Large-Scale Mutations
While point mutations are changes in a single nucleotide in...

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Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors
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Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors

Published on: September 20, 2016

iTILLING: personalized mutation screening.

Susan M Bush1, Patrick J Krysan

  • 1Department of Plant Biology, University of California-Davis, Davis, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 24, 2013
PubMed
Summary
This summary is machine-generated.

The iTILLING method offers a cost-effective and time-saving alternative to traditional TILLING for gene function studies. This approach enables individual labs to perform custom mutation screens in specific genetic backgrounds without establishing large mutant populations.

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Area of Science:

  • Plant science
  • Genetics
  • Molecular biology

Background:

  • Studying gene function often involves analyzing mutant phenotypes.
  • Efficiently isolating organisms with specific mutations is crucial for this research.
  • Traditional TILLING (Targeting Induced Local Lesions IN Genomes) relies on established, ordered mutant populations.

Purpose of the Study:

  • To introduce and describe the iTILLING method as an individualized approach to mutation screening.
  • To present iTILLING as a low-investment, practical alternative for individual research labs.
  • To highlight iTILLING's utility for custom mutagenesis screens in specific genetic backgrounds.

Main Methods:

  • iTILLING utilizes an ephemeral mutant population generated from Arabidopsis seedlings grown in 96-well plates.
  • It employs PCR and heteroduplex detection for mutation screening.
  • Unlike traditional TILLING, it does not require the creation of a durable, organized mutant population.

Main Results:

  • iTILLING significantly reduces the time and financial investment compared to traditional TILLING.
  • It provides a practical solution for mutation screening without the need for extensive resources.
  • This method allows for custom screens tailored to the specific research interests of individual labs.

Conclusions:

  • iTILLING is a valuable strategy for individual laboratories seeking to conduct targeted mutation screens.
  • It complements traditional TILLING by offering a flexible and resource-efficient option.
  • The method facilitates the study of gene function in unique genetic backgrounds relevant to specific research questions.