From Design to Single-Cell Cloning: A Complete RNP-Based CRISPR-Cas9 Protocol for Precision Gene Correction in Human iPSCs

Q: What does this research mean for the field?

An integrated RNP-based CRISPR-Cas9 protocol provides an efficient and comprehensive workflow for generating, screening, and expanding high-quality monoclonal human iPSC lines. Novelty: ClaimNovelty.METHODOLOGICAL. Consensus alignment: ConsensusAlignment.NEUTRAL.

Giacomo Roman; Knut H. Lauritzen; Barbora Smolkova; Runar Almaas; Per Morten Sandset; Benedicte Stavik; Gareth J. Sullivan; Maria E. Chollet

doi:10.1007/s12015-026-11221-6

August 31, 2026Stem Cell Reviews and ReportsOpen Access

From Design to Single-Cell Cloning: A Complete RNP-Based CRISPR-Cas9 Protocol for Precision Gene Correction in Human iPSCs

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Key Points

To establish a comprehensive, standardized workflow for editing, screening, and expanding monoclonal human induced pluripotent stem cell lines using ribonucleoprotein-based CRISPR-Cas9.
Outlines an integrated pipeline utilizing ribonucleoprotein (RNP) complexes of Cas9 and guide RNA for precision gene targeting.
Details single-cell deposition, high-throughput screening assays, and clonal expansion procedures for monoclonal line validation.
Provides a validated end-to-end framework that simplifies the isolation of pure, single-cell-derived edited colonies.
Ensures the maintenance of pluripotency and genomic integrity across the generation and expansion of edited human iPSC lines.

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Implication

Laboratory protocol demonstrates an integrated workflow for gene editing in human iPSCs, highlighting efficient generation of high-quality monoclonal lines.

Abstract

Summary This integrated end-to-end protocol provides a comprehensive and efficient workflow for the generation, screening and expansion of high-quality monoclonal iPSC lines after CRISPR-Cas9 editing.

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Roman et al. (2026) studied this question.

synapsesocial.com/papers/6a97e2b1c562ede874ec6fdf https://doi.org/10.1007/s12015-026-11221-6

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From Design to Single-Cell Cloning: A Complete RNP-Based CRISPR-Cas9 Protocol for Precision Gene Correction in Human iPSCs

Key Points

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Implication

Abstract

Discussion

Cite This Study

From Design to Single-Cell Cloning: A Complete RNP-Based CRISPR-Cas9 Protocol for Precision Gene Correction in Human iPSCs

Key Points

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Implication

Abstract

Discussion

Cite This Study

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