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Updated: May 28, 2025
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Delivery of the Cas9/sgRNA Ribonucleoprotein Complex in Immortalized and Primary Cells via Virus-like Particles "Nanoblades"
Published on: March 31, 2021
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Nano-Polymers as Cas9 Inhibitors.
Oksana Chepurna1, Avradip Chatterjee2, Yuanqing Li3,4
1Department of Neurosurgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Polymers
|February 13, 2025
Summary
Researchers found that biocompatible polymers like polymalic acid (PMLA), when linked to trileucine (LLL), can inhibit CRISPR-Cas9 gene editing. This discovery offers a new way to control and deliver CRISPR-Cas9 systems for precise gene editing applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Nanotechnology
Background:
- CRISPR/Cas9 technology is widely used but lacks effective delivery and control methods.
- Developing nanoscale delivery platforms with functional control is crucial for advancing gene editing.
Purpose of the Study:
- To develop a nanoscale CRISPR/Cas9 delivery platform with functional control.
- To identify biocompatible polymers that can inhibit Cas9 nuclease function.
Main Methods:
- Conjugation of biocompatible polymers (PMLA, PGA, PLD) with a trileucine (LLL) moiety.
- Assessing Cas9 nuclease inhibition by the polymer conjugates.
- Biophysical studies including protein binding assays.
- Transmission electron microscopy and molecular docking for mechanism elucidation.
Main Results:
- Several polymers, notably PMLA-LLL, effectively inhibited Cas9 nuclease activity in a dose-dependent manner.
- PMLA-LLL directly binds to the Cas9 protein, reducing Cas9/sgRNA binding affinity.
- A potential binding mechanism between PMLA-LLL and Cas9 was proposed via microscopy and docking.
Conclusions:
- A new class of Cas9 inhibitors in nano-polymer form has been identified.
- Biodegradable polymers like PMLA-LLL can serve as novel Cas9 delivery vehicles.
- These findings may enhance the precision of CRISPR-Cas9-mediated gene editing.
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