Cas12a

From WikiMD's Food, Medicine & Wellness Encyclopedia

Acidaminococcus sp. Cas12a PDB 5B43

Cas12a (also known as Cpf1) is a type of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) associated protein that is used in the field of genome editing. Cas12a is an endonuclease enzyme that is guided by a CRISPR RNA (crRNA) to target and cleave specific DNA sequences.

Discovery and Function[edit | edit source]

Cas12a was discovered as part of the CRISPR-Cas system in certain bacteria and archaea. Unlike the more commonly known Cas9 protein, Cas12a has unique properties that make it advantageous for certain genome editing applications. Cas12a recognizes a different protospacer adjacent motif (PAM) sequence, which is typically a T-rich sequence, and it creates staggered cuts in the DNA, leaving sticky ends.

Mechanism of Action[edit | edit source]

The Cas12a protein is guided to its target DNA by a crRNA. Upon binding to the target DNA, Cas12a induces a double-strand break. This break can be repaired by the cell's natural DNA repair mechanisms, such as non-homologous end joining (NHEJ) or homology-directed repair (HDR), leading to targeted modifications in the genome.

Applications[edit | edit source]

Cas12a has been utilized in various biotechnology and medical research applications, including:

  • Gene editing: Cas12a is used to introduce specific mutations or corrections in the genome.
  • Gene therapy: Potential use in treating genetic disorders by correcting defective genes.
  • Functional genomics: Studying gene function by creating targeted gene knockouts.

Advantages over Cas9[edit | edit source]

Cas12a offers several advantages over Cas9, including:

  • Simpler crRNA structure: Cas12a requires only a single crRNA, whereas Cas9 requires both a crRNA and a trans-activating crRNA (tracrRNA).
  • Different PAM requirements: Cas12a recognizes T-rich PAM sequences, expanding the range of targetable sites in the genome.
  • Staggered DNA cuts: The sticky ends created by Cas12a can facilitate more precise DNA insertions.

Challenges and Considerations[edit | edit source]

Despite its advantages, there are challenges associated with the use of Cas12a, such as off-target effects and delivery methods. Ongoing research aims to improve the specificity and efficiency of Cas12a-based genome editing.

See Also[edit | edit source]

References[edit | edit source]

External Links[edit | edit source]


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Contributors: Prab R. Tumpati, MD