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Using CRISPR-Kill for organ specific cell elimination by cleavage of tandem repeats

Angelina Schindele1, Fabienne Gehrke1, Carla Schmidt1

  • 1Botanical Institute - Molecular Biology and Biochemistry of Plants, Karlsruhe Institute of Technology, Fritz-Haber-Weg 4, 76131, Karlsruhe, Germany.

Nature Communications
|March 22, 2022
PubMed

Insights

CRISPR-Kill technology uses SaCas9 to induce targeted DNA breaks in repetitive regions, enabling controlled plant cell death for tissue engineering. This method allows precise manipulation of plant development, such as eliminating petals or reducing root growth.

Area of Science:

  • Plant Biology
  • Molecular Biology
  • Biotechnology

Background:

  • CRISPR/Cas systems are primarily used for gene editing via double-strand breaks (DSBs) in protein-coding genes.
  • Previous applications focused on gene knockout rather than controlled cell death induction.

Purpose of the Study:

  • To explore the use of CRISPR/Cas for controlled cell death induction in plants.
  • To develop CRISPR-Kill as a novel tool for plant tissue engineering and developmental manipulation.

Main Methods:

  • Utilized the SaCas9 nuclease to induce multiple DSBs in functional repetitive DNA of Arabidopsis thaliana.
  • Employed cell type-specific promoters to control the expression of SaCas9 and CRISPR-Kill.
  • Investigated the effects of CRISPR-Kill on organogenesis, petal development, and root architecture.

Main Results:

  • Demonstrated controlled induction of cell death in Arabidopsis thaliana by targeting repetitive DNA.
  • Successfully restored the apetala1 phenotype and eliminated petals by AP1-specific CRISPR-Kill expression.
  • Significantly reduced the number and length of lateral roots through root-specific pericycle cell expression of CRISPR-Kill.

Conclusions:

  • CRISPR-Kill is a viable tool for inducing controlled cell death and manipulating plant development.
  • This technology offers precise control over organogenesis and plant architecture.
  • Future applications may extend to altering plant biochemical properties.

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