Engineered PsCas9 enables therapeutic genome editing in mouse liver with lipid nanoparticles

Nov 7, 2024Nature communications

Engineered PsCas9 allows gene editing treatment in mouse liver using fat-based nanoparticles

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Abstract

Engineered PsCas9 (ePsCas9) demonstrates up to 20-fold increased editing activity compared to its predecessor.

  • PsCas9 is a Type II-B enzyme capable of editing the mouse liver genome with adenoviral delivery, showing no detectable off-target effects.
  • Non-viral delivery of PsCas9 remains a challenge due to insufficient efficacy.
  • Ribonucleoprotein structure of PsCas9 was solved using cryo-electron microscopy, aiding its rational engineering.
  • Multiple guide RNA and protein variants were screened to develop ePsCas9, which retains safety advantages while enhancing activity.
  • A single administration of mRNA encoding ePsCas9 with lipid nanoparticles achieved high editing levels in the Pcsk9 gene, relevant for hypercholesterolemia treatment.

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Key numbers

20×
Increase in Editing Activity
ePsCas9 activity compared to wild-type PsCas9 across various targets.
60%
Editing Efficiency in Mouse Liver
Average editing efficiency in the Pcsk9 gene after ePsCas9 delivery.
10×
Off-Target Activity Reduction
ePsCas9 off-target activity compared to SpCas9.

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