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ErCas12a CRISPR-MAD7 for Model Generation in Human Cells, Mice, and Rats.

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Summary

The MAD7 CRISPR-Cas system, derived from Eubacterium rectale, enables precise gene editing. It effectively creates genetic modifications and transgenic animals in various model systems.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas systems are powerful tools for genome engineering.
  • MAD7 is a class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) system from Eubacterium rectale.
  • Existing CRISPR systems have limitations in certain applications.

Purpose of the Study:

  • To evaluate the gene editing capabilities of the MAD7 system in human and animal cells.
  • To assess MAD7's efficiency in generating indels and performing gene tagging.
  • To determine MAD7's potential for creating transgenic animals.

Main Methods:

  • MAD7 system delivery into human HCT116 and U2OS cancer cell lines.
  • Application of MAD7 in mouse and rat embryos for generating transgenic animals.
  • Analysis of gene editing outcomes, including indels and insertions.

Main Results:

  • MAD7 successfully generated indels and fluorescent gene tagging in human cell lines.
  • MAD7 demonstrated high proficiency in creating indels and DNA insertions (up to 14 kb) in rodent embryos.
  • Live-born transgenic animals were produced using the MAD7 system.

Conclusions:

  • MAD7 is a versatile CRISPR-Cas tool with demonstrated gene editing activity in diverse cell types and organisms.
  • Its unique protospacer adjacent motif and small-guide RNA offer advantages over existing systems.
  • MAD7 expands the CRISPR toolbox for genome engineering in various research applications and model organisms.