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Point mutation KI cells
Cell line generation

Point mutation KI cells

A point mutation Knockin cell line has one or more mutated nucleotides that alter the protein amino acid sequence, changing its functions or properties.

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A point mutation Knockin cell line defines a model in which one or more nucleotides are constitutively mutated.

The insertion, deletion, nonsense and sense mutations alter the amino acid sequence of a given protein, thus changing its functions or properties.

Applications

For academic research:

  • Decipher protein function (gain or loss of function)‍
  • In vitro recapitulation of a human disease

For bio-pharmaceutical research & development:

  • Pharmacological off-target and efficacy studies by suppressing xenobiotics or antibody binding
  • Study drug-resistant mutants
  • Synthetic mortality screening in cancers

Strengths of point mutation Knockin cell models

  • High physiological relevancy of the scientific data obtained from the model (regulatory elements conserved, under control of endogenous promoter, expression of all splice variants, etc.) = more relevant than classical KO where the protein is absent
  • Reliable since each cell line is matched with an isogenic control cell clone

Limitations of point mutation Knockin cell models

  • Unpredicted alteration of splicing or 3D-protein structure

    →  If the genetic modification creates a dominant mutation, an overexpression of the point mutant in a safe harbor site (Quick Knockin), might be a valuable option

Validation

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Point mutation KI cells

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Inducible Knockdown cells

Through a single genetic modification, the addition of a self-cleaving degron tag to a protein of interest allows its inducible, and reversible, degradation upon treatment.

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Reporter KI cells

A reporter Knockin cell line is a useful model when there are no available antibodies for the target or when protein measurement is laborious and costly.

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Quick KI cells

A quick Knockin cell line has a transgene inserted into permissive loci (e.g., Rosa26). The transgene overexpression is constitutively or inducibly.

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Constitutive KO cells

A constitutive Knockout cell line refers to a conventional KO; a model in which the target gene is permanently inactivated, and no protein is produced.

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Point mutation Knockin cell lines

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