Cells undergo dramatic architectural remodeling in response to intrinsic and extrinsic perturbations. To capture a global picture of these changes under multiple conditions, we have developed the CZ Biohub “Organelle Box”, a panel of curated cell lines derived from A549 or HEK293, each labeled with an endogenous GFP-3XHA tag on specific organelle markers. This resource will facilitate the study of organelle dynamics, interactions, and functions across numerous models and research areas.

The available sets include:

• 21 A549 derived cell lines (Pilot distribution Mid 2025), representing 16 subcellular compartments

• 38 HEK293 derived cell lines (Pilot distribution Late 2025), representing 19 subcellular compartments

These panels of cell lines will be shared as an open access resource. For our pilot distribution, we welcome researchers to submit one paragraph proposals detailing how they plan to utilize these cells in their work.

We aim to distribute our pilot boxes to researchers who can:

• Maximize the use of the available cell lines (A549 or HEK293)

• Generate results within 6-12 months from delivery

• Apply diverse experimental perturbations (viral studies are of particular interest)

Researchers are encouraged to use and modify these cell lines for their studies and to acknowledge their use in scientific presentations and publications. We request that users share their observations with CZ Biohub San Francisco, which will allow us to compile and make these data available to the scientific community. These findings may also be included in consortium publications with appropriate attribution to all contributors.

• Who can get these cells?
  We invite investigators from diverse fields to consider using these cells in their studies. Our limited supply pilot boxes will be prioritized for researchers who can maximize using multiple cell lines within the next 6 months to 1 year.

• I want only one cell line, how can I get it?
  Unfortunately, we will only distribute the organelle box as a complete set. We cannot provide individual cell lines. We recommend that you contact other investigators who are currently using the box to learn more about potential collaborations or sharing.

• I do not see the marker I want in the box. Can you make another cell line?
  At this moment, we are not planning to expand our list of labeled targets. However, we encourage you to try labeling the proteins you're interested in yourself. We've included a protocol for cell engineering and can provide consultation if you encounter any troubleshooting challenges.

• What methods can I use in these cells?
  We welcome innovative and creative approaches! While we are currently performing organelle immunoprecipitation (Organelle-IP) and microscopy-based projects, we are genuinely excited to see the diverse and multiple ways other investigators might utilize these cells. Feel free to explore and innovate.

Please reach out to organellebox@czbiohub.org with your questions.