Structure-Guided Engineering of Compact Cas13d Ribonucleoproteins for High-Fidelity Transcriptome Editing

Chidi Okafor

doi:10.5555/peerrxiv.2026.00431.v1

PeerRxiv:2610.00431v1 · BIO-LIFE / Genomics & Gene Editing

Structure-Guided Engineering of Compact Cas13d Ribonucleoproteins for High-Fidelity Transcriptome Editing

Dr. Chidi Okafor (University of Cambridge)
ORCID: 0000-0002-6610-4982 · Pooled Credits: 0 Cr

CANONICAL CITATION ID

PeerRxiv:2610.00431v1

PEERRXIV VERSION DOI

10.5555/peerrxiv.2026.00431.v1

FREE CERN ZENODO DATACITE DOI

10.5281/zenodo.1400431

BAYESIAN SCORE / WILSON 95% LB

0.00 / 5.00 (LB 0.0%)

Abstract

Collateral trans-cleavage of bystander host transcripts limits the clinical translation of RNA-targeting CRISPR-Cas13 effectors. Using cryo-EM structure-guided mutagenesis of the HEPN1–HEPN2 catalytic cleft, we engineer a high-fidelity variant (hfCas13d-Max) that eliminates detectable transcriptome-wide collateral cleavage while preserving >88% on-target knockdown in human iPSC-derived cardiomyocytes.

Version Lineage (Before v1 vs. After v2)

V1 — Normal Preprint Archive (v1) · DOI: 10.5555/peerrxiv.2026.00431.v1 · 2026-10-27

Route 1 Normal Preprint Archive — Permanent Crossref DOI minted immediately with 0 credits required.

Public Signed Referee Reports (0)

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BibTeX Citation Record (Includes Signed Open Peer Reviews)

@article{okafor2026peerrxiv00431v1,
  author        = {Chidi Okafor},
  title         = {{Structure-Guided Engineering of Compact Cas13d Ribonucleoproteins for High-Fidelity Transcriptome Editing}},
  journal       = {PeerRxiv Open Science Repository},
  year          = {2026},
  month         = oct,
  eid           = {PeerRxiv:2610.00431v1},
  eprint        = {PeerRxiv:2610.00431v1},
  archivePrefix = {PeerRxiv},
  primaryClass  = {bio-life},
  issn          = {0000-0000},
  doi           = {10.5555/peerrxiv.2026.00431.v1},
  note          = {PeerRxiv Open Preprint (NORMAL PREPRINT); Dual Free DataCite DOI: 10.5281/zenodo.1400431},
  url           = {http://peerrxiv.org/abs/peerrxiv.2026.00431}
}