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pegRNA

Definition

pegRNA, or prime editing guide RNA, is a specialized RNA molecule used in prime editing, a next-generation genome-editing technology. Prime editing offers a more precise and flexible method for introducing specific genetic changes, including base insertions, deletions, and substitutions, without creating double-stranded breaks in the DNA, as is the case with CRISPR-Cas9. The pegRNA guides the prime editor to the target DNA sequence and also encodes the desired edit.

Structure

pegRNA consists of two key components:

  • Spacer Region: This acts like a conventional guide RNA (gRNA) in CRISPR systems, guiding the prime editor to the specific target sequence in the genome.
  • Reverse Transcriptase Template: This region contains the sequence of the desired edit and serves as a template for the reverse transcriptase component of the prime editor to copy the modification directly into the genome.

Function

In prime editing, pegRNA works alongside a prime editor enzyme, which is a fusion of a Cas9 nickase (a mutated form of Cas9 that only cuts one strand of DNA) and a reverse transcriptase. The pegRNA directs the prime editor to the target site, where the reverse transcriptase uses the pegRNA’s template to synthesize the new DNA strand, introducing the intended edit with high precision.

Applications

Prime editing, utilizing pegRNA, is a valuable technique for precise genome engineering. It is used in:

  • Correcting point mutations associated with genetic diseases.
  • Introducing targeted modifications without relying on donor DNA templates or causing double-stranded breaks.
  • Creating or repairing mutations in research models, offering more controlled editing outcomes compared to earlier genome-editing technologies.

Advantages

  • Precision: Prime editing, guided by pegRNA, enables highly specific edits with fewer off-target effects.
  • Versatility: It can introduce a wide range of genetic modifications (insertions, deletions, base substitutions).
  • Reduced Damage: Unlike traditional CRISPR-Cas9, pegRNA and prime editing do not rely on double-strand breaks, minimizing potential genomic instability.

The innovative tool of pegRNA is expanding the possibilities for genetic research, therapeutic development, and synthetic biology.


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