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SpRY: Engineered CRISPR/Cas9 Harnesses New Genome-Editing Power
Dangquan Zhang1, Baohong Zhang2
1Henan Province Engineering Research Center for Forest Biomass Value-Added Products, College of Forestry, Henan Agricultural University, Zhengzhou, Henan 450002, China.
Trends in Genetics : TIG
|May 28, 2020
Summary
CRISPR/Cas9 gene editing is limited by PAM requirements. Researchers engineered a near-Protospacer Adjacent Motif (PAM)-less SpRY variant of Streptococcus pyogenes Cas9 (SpCas9) that efficiently targets most DNA sequences, expanding genome-editing applications.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- CRISPR/Cas9 gene editing technology relies on Protospacer Adjacent Motif (PAM) sequences for DNA targeting.
- The PAM requirement restricts CRISPR/Cas9 accessibility to numerous genetic loci, limiting its research and therapeutic potential.
Purpose of the Study:
- To overcome the PAM limitations of CRISPR/Cas9.
- To engineer a novel Cas9 variant with expanded DNA targeting capabilities.
Main Methods:
- Structural engineering of Streptococcus pyogenes Cas9 (SpCas9).
- Development of a near-Protospacer Adjacent Motif (PAM)-less SpRY variant.
Main Results:
- The engineered SpRY variant demonstrates the ability to target most DNA sequences.
- High editing efficiency and flexibility were observed with the SpRY system.
- The SpRY system overcomes the PAM specificity of conventional SpCas9.
Conclusions:
- The near-PAMless SpRY variant significantly broadens the scope of CRISPR-based genome editing.
- This advancement holds potential for diverse applications in basic and applied biological research.
- SpRY offers enhanced flexibility and efficiency for precise genetic modifications.
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