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Immortalised Cas9-expressing Cell lines for Gene interrogation.

Luis F Malaver-Ortega1,2, Joseph Rosenbluh3,4

  • 1Monash Functional Genomics Platform, Monash University, Clayton, VIC, Australia. LuisMalaver@gmail.com.

Methods in Molecular Biology (Clifton, N.J.)
|June 13, 2022
PubMed
Summary

This study presents a practical pipeline for creating immortalized cell lines from various tissues. This method aids in validating genome editing tools and understanding gene function across species.

Keywords:
CRISPR/Cas9Cas9 assayCell immortalizationGenome editing

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Area of Science:

  • Biotechnology and Genetic Engineering
  • Cell Biology and Tissue Engineering

Background:

  • Precise genome modification with minimal off-target effects is crucial for biotechnology.
  • The development of robust cell line generation methods is needed for genetic research.

Purpose of the Study:

  • To describe a practical pipeline for generating immortalized cell lines.
  • To provide a method for validating genome editing tools and genetic modifications.
  • To establish a reference for gene interrogation in diverse species.

Main Methods:

  • Development of an easy-to-establish, robust, and practical pipeline.
  • Generation of immortalized cell lines from different tissue types.
  • Utilizing cell lineage context for tool validation.

Main Results:

  • Successful generation of immortalized cell lines from various tissues.
  • Demonstration of the pipeline's utility in capturing cell lineage context.
  • Validation of genome editing and genetic modification tools.

Conclusions:

  • The described pipeline offers a reliable method for generating immortalized cell lines.
  • This approach facilitates the validation of genome editing technologies.
  • The pipeline serves as a valuable reference for cross-species gene interrogation.