An Overview and Comparative Analysis of CRISPR-SpCas9 gRNA Activity Prediction Tools

CRISPR J. 2025 Apr;8(2):89-104. doi: 10.1089/crispr.2024.0058. Epub 2025 Mar 27.

Abstract

Design of guide RNA (gRNA) with high efficiency and specificity is vital for successful application of the CRISPR gene editing technology. Although many machine learning (ML) and deep learning (DL)-based tools have been developed to predict gRNA activities, a systematic and unbiased evaluation of their predictive performance is still needed. Here, we provide a brief overview of in silico tools for CRISPR design and assess the CRISPR datasets and statistical metrics used for evaluating model performance. We benchmark seven ML and DL-based CRISPR-Cas9 editing efficiency prediction tools across nine CRISPR datasets covering six cell types and three species. The DL models CRISPRon and DeepHF outperform the other models exhibiting greater accuracy and higher Spearman correlation coefficient across multiple datasets. We compile all CRISPR datasets and in silico prediction tools into a GuideNet resource web portal, aiming to facilitate and streamline the sharing of CRISPR datasets. Furthermore, we summarize features affecting CRISPR gene editing activity, providing important insights into model performance and the further development of more accurate CRISPR prediction models.

Publication types

  • Review

MeSH terms

  • Animals
  • CRISPR-Associated Protein 9* / genetics
  • CRISPR-Associated Protein 9* / metabolism
  • CRISPR-Cas Systems* / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats* / genetics
  • Computational Biology / methods
  • Computer Simulation
  • Deep Learning
  • Gene Editing* / methods
  • Humans
  • Machine Learning
  • RNA, Guide, CRISPR-Cas Systems* / genetics
  • Software

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • CRISPR-Associated Protein 9