Applied StemCell Demonstrates Efficient Knock-In of a 50 kb DNA Construct into Human iPSCs Using TARGATT™ Technology

MILPITAS, Calif., Nov. 11, 2025 (GLOBE NEWSWIRE) -- Applied StemCell, Inc. (ASC), today announced the successful knock-in of a 50 kilobase (kb) multifunctional DNA insert into human induced pluripotent stem cells (iPSCs) using its proprietary TARGATT™ technology. The achievement establishes a new benchmark for the size and complexity of genetic payloads that can be efficiently and site-specifically integrated into the human genome.

The 50 kb DNA construct — produced by Ansa Biotechnologies using its enzymatic DNA synthesis platform — was integrated into the H11 safe harbor locus of human iPSCs with greater than 20% efficiency and delivered robust, consistent expression of multiple genes. This work represents a significant advance in the ability to perform large, precise, and reproducible genome engineering in human cells.

“This milestone demonstrates that TARGATT™ technology can efficiently integrate DNA constructs more than twice as large as previously validated payloads,” said Dolores Baksh, Ph.D., CEO of Applied StemCell. “This opens the door to a wide range of applications that demand stable, site-specific insertion and consistent gene expression, from complex protein expression systems to next-generation allogeneic cell therapies.”

The 50 kb construct contains multiple expression cassettes — including constitutive and inducible genes — enabling both immune evasion and tunable gene expression. Functional assays confirmed targeted integration and expression fidelity across the edited iPSC population, with virtually 100% of integrated cells expressing a constitutive transgene and inducible genes responding predictably to doxycycline treatment.

This capability unlocks new possibilities for:

• Allogeneic cell therapy development, enabling the creation of immune-evasive or hypoimmunogenic cell lines.
• Multi-protein complex expression, where precise stoichiometry is essential.
• Stable packaging and production lines for viral vector and protein manufacturing.

The success of this project also underscores the value of Ansa Biotechnologies’ ultra-long DNA synthesis technology, which allowed the full 50 kb donor plasmid to be built in less than one month — significantly faster than conventional cloning approaches that can take several months.

“Constructs of this size are notoriously difficult to build using standard cloning methods,” said Daniel Lin-Arlow, Ph.D., CSO and co-founder of Ansa Biotechnologies. “Our enzymatic synthesis and assembly technologies eliminate that bottleneck, allowing Applied StemCell to focus on using their TARGATT technology for cell engineering instead of troubleshooting donor plasmid construction.”

Together, these advances demonstrate how Ansa’s rapid DNA synthesis and Applied StemCell’s efficient, site-specific knock-in technologies can accelerate the development of complex engineered mammalian cells for research and therapeutic applications.

About Applied StemCell, Inc.
Applied StemCell (ASC) is a genome engineering platform company advancing the future of genetic and cell-based medicine through its proprietary TARGATT™ and S-SELeCT™ technologies. These innovations enable safe, scalable, site-specific integration of large payloads with reliable gene expression for the development of next-generation therapies, ex vivo and in vivo.

With deep expertise in iPSCs and cell line engineering, ASC provides end-to-end solutions that accelerate innovation across academia and biopharma. Our strong IP foundation and cutting-edge technologies empower partners to move from concept to clinic with precision, scalability, and confidence. www.appliedstemcell.com

For media inquiries, please contact:
Pia Abola, Ph.D.
Director of Marketing
Applied StemCell
pia.abola@appliedstemcell.com
(408) 457-1312 

A photo accompanying this announcement is available at https://www.globenewswire.com/NewsRoom/AttachmentNg/53018311-088c-4f9f-950e-62de140420df