ImmunityJul 9, 2026

Using CRISPR to quickly add large human antibody genes into mouse embryos

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jul 10, 2026

Abstract

A CRISPR-guided approach allows the insertion of a 155-kb human genomic sequence into mouse zygotes.

Large genomic edits can be achieved by deleting a 2.4-Mb mouse immunoglobulin heavy-chain locus.
A bacterial artificial chromosome containing human DNA was successfully integrated without unwanted recombination.
The inserted human sequences were stably passed on and expressed, interacting with mouse genetic material.
Mice developed normal B cells and demonstrated class-switch recombination and somatic hypermutation after immunization.
The method produces genetically modified mice with large insertions in approximately 8 weeks, significantly faster than traditional techniques.

Simplified

Current CRISPR-Cas9 methods are restricted to small genomic edits. We developed a CRISPR-guided approach that enables direct insertion of large genomic sequences into mouse zygotes. We deleted the murine 2.4-Mb immunoglobulin heavy-chain (IgH) variable (V_H) locus and then precisely inserted a bacterial artificial chromosome (BAC) containing a 155-kb human V_H DNA fragment flanked by 20-kb homology arms. Full-length, single-copy BAC integration occurred without ectopic recombination. Human sequences were stably transmitted and expressed V_H segments that recombined with endogenous mouse sequences, and mice exhibited normal B cell development. Upon immunization, human V_H-expressing B cells underwent class-switch recombination and somatic hypermutation, secreting antigen-specific antibodies. We demonstrated modular IgH humanization by replacing endogenous mouse diversity and joining (D_H-J_H) segments with human sequences, producing V(D)J recombination and diverse antibodies. Unlike traditional methods requiring more than a year, this approach enables the generation and validation of mice carrying large genomic insertions within 8 weeks.

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Competing interests

Declaration of interests U.N. and F.D.B. have filed a provisional patent for the described genome modification methods. F.D.B. has consultancy relationships with Adimab, Third Rock Ventures, and The EMBO Journal and also founded BliNK Therapeutics. D.L. reports consultancy relationships with Metaphore Bio (a Flagship company), Tendel Therapies, and BioMed X. D.L.’s laboratory has also received funding from Leyden Labs for unrelated work.

PubMed

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CRISPR Gene EditingIssue #45

Scientists built humanized antibody mice in 8 weeks instead of over a year

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Using CRISPR to quickly add large human antibody genes into mouse embryos

Abstract

Full Text

Funding

Competing interests

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