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Applied StemCell, TARGATT™

Site-Specific Knock-in Technology

High integration efficiency

Up to 20 kb Transgene Knock-In

Fast Turnaround

Reasons to Consider TARGATT™ Genome Engineering Technology for Safe and Efficacious Induced Pluripotent Stem Cell Therapies

  1. Specificity & Safety: unidirectional site-specific integration into a preselected locus
  2. Gene Editing Efficiency: high-efficiency site-specific insertion of large transgenes, achieving insertion efficiencies as high as 40% in many cell lines, including iPSCs
  3. Editing In Non-Dividing Cell Lines: facilitates targeted genetic modifications in quiescent and/or terminally differentiated cells, unlocking novel opportunities for disease modeling, drug discovery, target discovery, and tissue regeneration
  4. Payload Capacity: integration of transgenes of up to 20 kb 
  5. Manufacturing & Outsourcing: avoiding the complexities associated with viral vector production with FTO for commercialization
Want To Learn More About iPSC Applications? Case Studies Are Available

TARGATT™ gene editing technology is a highly efficient gene editing platform. Its foundational technology was co-invented by Applied StemCell’s CEO, Dr. Chen-Tsai and R&D Head, Dr. Farruggio while they were at Stanford University. ASC’s research team has been developing and improving the technology. 

TARGATT™gene editing technology facilitates rapid and precise integration of large DNA fragments (up to 20 kb) into a specific intergenic locus. Single-copy gene integration is achieved at the preselected locus by utilizing an “attP” integrase recognition landing pad in conjunction with an “attB” containing donor plasmid and integrase expression. This unique combination ensures stable integration into a transcriptionally active safe harbor site with exceptional efficiency. TARGATT™ is an excellent tool for the development of allogenic Induced Pluripotent Stem Cell (iPSC) therapies and advancing research in regenerative medicine, immune cell therapy, and genomics.  

The TARGATT™ gene editing platform is incredibly versatile, making it ideal for creating large fragment knock-in cell lines, protein expression, and library construction. This technology represents a significant advancement in gene editing by circumventing issues traditionally associated with random integration, such as position effect, gene silencing, and gene instability due to the integration of multiple transgene copies.

Ask an Expert About TARGATT™

TARGATT™ Gene Editing Compared To Traditional Gene Editing Technologies

To develop safe and effective cell therapies, it is important to have a comprehensive awareness of the benefits, limitations, and opportunities made possible by each gene editing platform on the market. Traditional gene insertion technologies like Lentivirus, Transposase, and even CRISPR have historically had limitations related to random insertion and off-target insertion risks.

targattApplied StemCell’s proprietary TARGATT™ genome engineering platform offers a compelling solution. It ensures specificity, efficiency, safety, and the ability to insert genes in a site-specific manner.  With a substantial payload capacity and a clear IP path to commercialization, TARGATT™ streamlines the manufacturing process, saving time and cost.

 By leveraging the unique advantages of TARGATT™, researchers can expedite the development of allogeneic iPSC-based cell therapies, paving the way for a new era of patient-centric treatment modalities.

  • Technical Details

    Figure 1: TARGATT™ Knock-In Strategy. TARGATT™ technology enables fast and site-specific, stable integration of large DNA fragments (up to 20 kb) into an intergenic, transcriptionally active safe harbor locus. We can engineer an “attP” integrase recognition landing pad at a safe harbor locus. Single-copy gene insertion occurs when it is used in conjunction with an “attB” containing donor plasmid and integrase expression.

Frequently Asked Questions

  • How does the TARGATT™ knock-in system achieve high efficiency?

    The TARGATT™ integrase system operates differently from other recombinases like Cre or Flp. It recognizes and recombines at two distinct sites, attP, and attB, which are largely unrelated in terms of their sequences. Upon integrase-mediated integration at attB and attP, two new hybrid sites, attL, and attR, are formed at the junctions. These hybrid sites cannot be recognized by the integrase, resulting in a unidirectional integrase reaction. Once the DNA is integrated, it remains permanently inserted at the designated locus such as AAVS1 or H11 locus without the possibility of excision. This characteristic ensures a highly efficient and stable integration process using the TARGATT™ integrase system.

     

  • Do you have TARGATT™ technology available for Knock-in cell lines?

    Yes, we offer TARGATT™ Knock-in ready cell lines in three cell types. They are HEK293, CHO and iPSC lines. We can also generate TARGATT™ master cell lines engineered with the “attP” docking site at the safe harbor locus in your cell line of interest.

  • When using TARGATT™, how many copies of the gene will be inserted into the genome?

    A single copy of each gene of interest will be inserted.

  • What promoters are used to drive gene expression?

    Any defined promoters provided by the customer or published in the literature can be used to drive gene expression.

  • What is the specific site that my gene of interest will be integrated into?

    If using our off-the-shelf TARGATT™ knockin-ready cells, your gene of interest will be specifically inserted at the H11 locus. We can also engineer the landing pad at the locus of your choice.

  • What is the maximum size of a gene you can insert? Will the efficiency of your system be affected if the gene is too large?

    To date, we have successfully integrated DNA fragments up to 20 kb. Efficiency may decrease with larger fragments.

  • Can I create models to overexpress a gene of interest?

    Yes, the TARGATT™ system is ideal for gene over-expression. Different promoters (e.g., tissue-specific promoters or ubiquitous promoters), and inducible systems (e.g., Tet On/Off, loxP-stop-loxP) can be used for tissue-specific, ubiquitous, or inducible gene expression.

  • Can I integrate a reporter gene using TARGATT™? What kind of reporter genes do you recommend?

    Yes, you can express any reporter genes using TARGATT™ technology. Common examples include, but are not limited to: GFP, DsRed, mCherry, LacZ, and Luciferase.

Contact us now to learn how TARGATT™ can benefit your research.

MTA Agreement

Material Transfer Agreement

IMPORTANT! PLEASE READ CAREFULLY BEFORE SUBMITTING AN ORDER. THIS IS A CONTRACT.

This Material Transfer Agreement (“MTA”) is between you (“Purchaser”) and the Applied StemCell, Inc. a California company, having its principal place of business at 521 Cottonwood Drive Suite 111, Milpitas, CA 95035 USA (“ASC”). Purchaser must have an approved, current ASC account to place an order. This MTA is effective for a period of five (5) years as of the last date of execution by the parties and governs the purchase and use of all ASC Materials under the terms and conditions set forth below.

TERMS AND CONDITIONS

Definitions

“ASC Material(s)” means materials acquired from ASC as documented on an ASC Sales Order, such as iPS Cell lines.

"ASC Sales Order" means an order submitted for ASC Materials in a form and format as determined by ASC from time to time. "Biological Material(s)" means ASC Materials, Progeny, Unmodified Derivatives and Modifications, either individually or jointly. "Commercial Use" means the sale, license, lease, export, transfer or other distribution of the Biological Materials to a person or entity not party to this MTA for financial gain or other commercial purposes and/or the use of the Biological Material: (a) to provide a service to a person or entity not party to this MTA for financial gain; (b) to produce or manufacture products for general sale or products for use in the manufacture of products ultimately intended for general sale (c) in connection with ADME (Absorption, Distribution, Metabolism and Excretion) testing; (d) in connection with drug potency or toxicity testing (e) in connection with proficiency testing service(s), including but not limited to, providing the service of determining laboratory performance by means of comparing and evaluating calibrations or tests on the same or similar items or materials in accordance with predetermined conditions; or (f) for research conducted under an agreement wherein a for-profit entity receives a right whether actual or contingent to the results of the research. Commercial Use specifically does not include Industry Sponsored Academic Research.

“Contributor(s)” means an organization(s) and/or individual(s) providing original material to ASC for deposit.

"Industry Sponsored Academic Research" means research sponsored by a for-profit organization carried out at a non-profit organization and by the non-profit organization’s employees. "Investigator" means the Purchaser’s principal scientist or researcher using the Biological Material(s). "Modification(s)" mean substances created by Purchaser which contain and/or incorporate a significant or substantial portion of ASC Material. "Progeny" means an unmodified descendant from the ASC Materials, such as virus from virus, cell from cell, or organism from organism. "Purchaser(s)" means the organization purchasing and receiving ASC Material pursuant to this MTA. "Unmodified Derivative(s)" mean substances created by Purchaser that constitute an unmodified functional sub-unit or product not changed in form or character and expressed by the ASC Material provided by ASC. Unmodified Derivatives include, but are not limited to, subclones of unmodified cell lines, purified or fractionated subsets of materials provided by ASC, proteins expressed by DNA/RNA supplied by ASC, or monoclonal antibodies secreted by a hybridoma cell line.

Scope of Use

Subject to the terms of this MTA, Purchaser’s Investigator may make and use the Biological Materials provided to Purchaser by ASC for research purposes only in Purchaser’s Investigator’s laboratory only. The Biological Materials are not intended for use in humans. Purchaser agrees that Biological Materials designated as biosafety level 2 or 3 constitute known pathogens and that other Biological Materials not so designated may be pathogenic under certain conditions. Purchaser assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, transfer and Purchaser’s use of the Biological Materials including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. Purchaser agrees that any activity undertaken with the Biological Materials will be conducted in compliance with all applicable guidelines, laws and regulations, and that Purchaser will obtain all permits, licenses or other approvals required by any governmental authority in connection with purchaser’s receipt, handling, storage, disposal, transfer and use of the Biological Materials.

Purchaser shall not distribute, sell, lend or otherwise transfer to a person or entity not party to this MTA the Biological Material, as defined above, for any reason, without ASC’s prior written agreement.

Any Commercial Use of the Biological Material is strictly prohibited without ASC’s prior written consent. Purchaser acknowledges and agrees that Purchaser’s use of certain Biological Material may require a license from a person or entity not party to this MTA, or be subject to restrictions that may be imposed by a person or entity not party to this MTA (“Third Party Terms”). To the extent of ASC’s knowledge of the existence of any such applicable rights or restrictions, ASC will take reasonable steps to identify the same, either in ASC’s catalog of ASC Materials and/or through ASC’s customer service representatives, and to the extent they are in the possession of ASC, ASC shall make information regarding such Third Party Terms reasonably available for review by Purchaser upon request. Purchaser expressly acknowledges that if there is a conflict between this MTA and the Third Party Terms, the Third Party Terms shall govern. Use of the Biological Materials may be subject to the intellectual property rights of a person or entity not party to this MTA, the existence of which rights may or may not be identified in the ASC catalog or website, and ASC makes no representation or warranty regarding the existence or the validity of such rights. Purchaser shall have the sole responsibility for obtaining any intellectual property licenses necessitated by its possession and use of the Biological Materials.

The use permitted under this MTA for Industry Sponsored Academic Research extends only to the academic research carried out at the non-profit organization and the non-profit organization’s employees. Any non-profit Purchaser using the Biological Materials in connection with Industry Sponsored Academic Research agrees to notify the industrial sponsor that any use of the Biological Materials by the industry sponsor will require a separate license from ASC and/or its Contributors and that ASC and/or its Contributors are under no obligation hereunder to license any Biological Materials to any such industry sponsor.

Warranty; Warranty Disclaimer

ASC warrants that (a) cells and microorganisms included in the ASC Material shall be viable upon initiation of culture for a period of thirty (30) days after shipment thereof from ASC and (b) any ASC Material other than cells and microorganisms shall meet the specifications on the applicable ASC Material product information sheet, certificate of analysis, and/or catalog description until the expiration date on the applicable ASC Material’s product label (such thirty (30) day period, or period until the expiration date, referred to herein as the “Warranty Period”). Purchaser’s exclusive remedy, and ASC’s sole liability, for breach of the warranties set forth in this paragraph is for ASC to, at ASC’s sole option, either (i) refund the fee paid to ASC for such ASC Material (exclusive of shipping and handling charges), or (ii) replace the ASC Material. The warranties set forth in this paragraph apply only if Purchaser handles and stores the ASC Material as described in the applicable ASC Material product information sheet. To obtain the exclusive remedy, Purchaser must report the lack of viability or non-conformation to specifications to ASC’s Technical Service Department within the applicable Warranty Period. Any expiration date specified on the ASC Material shipment documentation states the expected remaining useful life, but does not constitute a warranty or extend any applicable Warranty Period. Except as expressly provided above, the ASC Material and any technical information and assistance provided by ASC are provided as-is, without warranties of any kind, express or implied, including but not limited to any implied warranties of merchantability, fitness for a particular purpose, typicality, safety, accuracy and/or non-infringement.

Purchaser acknowledges that the ASC Material and any technical information and assistance provided by ASC are developed and provided exclusively for research purposes, and Purchaser agrees to use the same at its sole risk. Purchaser agrees that ASC and its Contributors will not be liable for any loss, claim or demand made by Purchaser, or made against Purchaser by any other party, due to or arising from the use of the ASC Material by Purchaser, except to the extent permitted by law when caused by the gross negligence or willful misconduct of ASC.

Limitation of Liability

In no event shall ASC or its Contributors be liable for any use of the Biological Material by Purchaser, for any loss, claim, damage, or liability, of whatever kind or nature, which may arise from or in connection with this MTA or the use, handling, storage, or disposal of the Biological Material. ASC’s liability to Purchaser for any claim related to or arising from this MTA or the Biological Material, whether in contract, warranty, tort, or otherwise, shall be limited to the amount paid by Purchaser for the Biological Material. In no event shall ASC be liable for any indirect, special, incidental, consequential, or punitive damages, including without limitation, loss of profits or loss of use, even if ASC has been advised of the possibility of such damages. The limitations of liability set forth in this MTA shall survive termination or expiration of this MTA for any reason.