CLASH enables large-scale parallel knock-in for cell engineering

    Nature Biotechnology

    (2023)Cite this article

    Subjects

    CLASH is a CRISPR-based platform that enables the parallel knock-in via homology-directed repair of a large pool of transgene variants encoded in adeno-associated virus vectors. CLASH can be applied to the systematic and unbiased selection of favorable features in T cells and, in principle, other cell types.

    This is a preview of subscription content, access via your institution

    Access options

    Subscribe to Nature+

    Get immediate online access to Nature and 55 other Nature journal

    Subscribe to Journal

    Get full journal access for 1 year

    $99.00

    only $8.25 per issue

    All prices are NET prices.
    VAT will be added later in the checkout.
    Tax calculation will be finalised during checkout.

    Buy article

    Get time limited or full article access on ReadCube.

    $32.00

    All prices are NET prices.

    Fig. 1: Development of CLASH system for massively parallel knock-in engineering and selection in primary human T cells.

    References

    1. Dong, M. B. et al. Systematic immunotherapy target discovery using genome-scale in vivo CRISPR screens in CD8 T sells. Cell 178, 1189–1204.e1123 (2019).

      Article 
      CAS 

      Google Scholar
       

    2. Long, L. et al. CRISPR screens unveil signal hubs for nutrient licensing of T cell immunity. Nature 600, 308–313 (2021).

      Article 
      CAS 

      Google Scholar
       

    3. Chen, Z. et al. In vivo CD8+ T cell CRISPR screening reveals control by Fli1 in infection and cancer. Cell 184, 1262–1280.e1222 (2021). Refs. 1–3 are representative lentivirus-mediated CRISPR screens in primary T cells.

      Article 
      CAS 

      Google Scholar
       

    4. Roth, T. L. et al. Pooled knockin targeting for genome engineering of cellular immunotherapies. Cell 181, 728–744.e21 (2020). Mini-pool knock-in in human primary T cells.

      Article 
      CAS 

      Google Scholar
       

    5. Schumann, K. et al. Generation of knock-in primary human T cells using Cas9 ribonucleoproteins. Proc. Natl Acad. Sci. USA 112, 10437–10442 (2015). CRISPR knock-in in human primary T cells.

      Article 
      CAS 

      Google Scholar
       

    Download references

    Additional information

    Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

    This is a summary of: Dai, X. et al. Massively parallel knock-in engineering of human T cells. Nat. Biotechnol. https://doi.org/10.1038/s41587-022-01639-x (2023).

    About this article

     Verify currency and authenticity via CrossMark

    Cite this article

    CLASH enables large-scale parallel knock-in for cell engineering.
    Nat Biotechnol (2023). https://doi.org/10.1038/s41587-023-01664-4

    Download citation

    • Published:

    • DOI: https://doi.org/10.1038/s41587-023-01664-4

    Read More
    Laine Byron

    Latest

    Green steel startup Boston Metal is doubling down on critical metals

    The startup Boston Metal has raised a $75 million funding round to produce critical metals, MIT Technology Review can exclusively report.   The company has been known largely for its efforts to clean up steel production, an industry that's responsible for about 8% of global greenhouse emissions today. With the additional money, the new focus could

    Embracer Follows Ubisoft In Splitting Off New Publisher To Handle Huge IP, Tomb Raider & LOTR Included

    Say hello to Fellowship Entertainment by Ben Kerry 11 hours ago Embracer Group has today announced plans to create a secondary publishing label called Fellowship Entertainment, in order to "capture the full potential of the high-quality assets" that the group currently owns. The Swedish game publisher says that it hopes to spin off Fellowship Entertainment

    Gwyneth Paltrow’s Daughter Apple Martin in Nancy Meyers Movie

    Gwyneth Paltrow's Daughter Apple Martin Makes Directorial Debut With Student Show Apple Martin doesn’t fall far from the tree. Gwyneth Paltrow and Chris Martin ’s daughter will be following in her mom’s acting footsteps and making her movie debut in Nancy Meyers’ upcoming film, Deadline and Entertainment Weekly reported on May 18. The 22-year-old—who graduated

    Newsletter

    Don't miss

    Green steel startup Boston Metal is doubling down on critical metals

    The startup Boston Metal has raised a $75 million funding round to produce critical metals, MIT Technology Review can exclusively report.   The company has been known largely for its efforts to clean up steel production, an industry that's responsible for about 8% of global greenhouse emissions today. With the additional money, the new focus could

    Embracer Follows Ubisoft In Splitting Off New Publisher To Handle Huge IP, Tomb Raider & LOTR Included

    Say hello to Fellowship Entertainment by Ben Kerry 11 hours ago Embracer Group has today announced plans to create a secondary publishing label called Fellowship Entertainment, in order to "capture the full potential of the high-quality assets" that the group currently owns. The Swedish game publisher says that it hopes to spin off Fellowship Entertainment

    Gwyneth Paltrow’s Daughter Apple Martin in Nancy Meyers Movie

    Gwyneth Paltrow's Daughter Apple Martin Makes Directorial Debut With Student Show Apple Martin doesn’t fall far from the tree. Gwyneth Paltrow and Chris Martin ’s daughter will be following in her mom’s acting footsteps and making her movie debut in Nancy Meyers’ upcoming film, Deadline and Entertainment Weekly reported on May 18. The 22-year-old—who graduated

    Lil Wayne speaks out after feeling overlooked by Coachella and the Grammys

    Music Lil Wayne reacts to Coachell and Grammys snub Award-winning...

    Tesla’s Business Has Become Much More Diversified in Just the Past Five Years. Does That Make Its Stock a Better Buy Today?

    Key Points Tesla's energy generation and storage segment generated 27% revenue growth last year. The company's non-automotive segments were able to help offset a double-digit decline in auto revenue in 2025. These 10 stocks could mint the next wave of millionaires › Tesla (NASDAQ: TSLA) is known for its electric vehicles (EVs), and while they

    WD sees sustainability as key business driver in an ‘AI economy’

    Hard drive company WD promoted long-term operations and sustainability executive Jackie Jung to become its first chief sustainability officer in February, as it steps up sales to companies building AI data centers. Her vision: Turn sustainability into a “brand” for WD, a strategy that reduces risk for the $6 billion company (formerly known as Western

    5 Business Ideas Worth Starting in 2026

    If there is one thing Nigerians understand well, it is how to spot opportunity inside hardship. In 2026, that mindset will matter more than ever. The economy is tough, competition is rising, and many people are looking for smarter ways to earn, build, and survive. But even in a difficult environment, some businesses still stand