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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Related Experiment Video

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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
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A Split CRISPR-Cpf1 Platform for Inducible Gene Activation.

Takahiro Otabe1,2, Yuta Nihongaki1,3, Moritoshi Sato4,5

  • 1Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|September 29, 2022
PubMed
Summary
This summary is machine-generated.

Researchers engineered photoactivatable CRISPR-Cpf1 (paCpf1) for precise, light-controlled genome editing. They also developed a split Cpf1 activator for endogenous gene activation in cells and animals.

Keywords:
CRISPR–Cpf1 (also known as Cas12a)Gene activationGenome editingGenomic PCRHydrodynamic tail vein injectionIn vivo gene activationNHEJOptogeneticsT7E1crRNA

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cpf1 (Cas12a) is an RNA-guided endonuclease.
  • Optogenetics enables light-based control of biological systems.

Purpose of the Study:

  • To develop optogenetic tools for precise genome editing.
  • To create a system for endogenous gene activation.

Main Methods:

  • Engineered photoactivatable Cpf1 (paCpf1) by combining CRISPR-Cpf1 with optogenetics.
  • Identified a spontaneously activated split Cpf1.
  • Developed a dCpf1 split activator for gene activation.

Main Results:

  • Demonstrated optogenetic endogenous genome editing using paCpf1 in mammalian cells.
  • Showcased endogenous gene activation using the dCpf1 split activator in mammalian cells and mice.

Conclusions:

  • paCpf1 allows spatiotemporal control of genome editing.
  • The dCpf1 split activator system offers a potent method for endogenous gene activation.