Lentiviruses are the backbone of many engineered cell therapies, but remain expensive to make and difficult to scale. Our LV Edge system can help. Part I of II.
Excellent article! I wonder what your team thinks about non-viral engineering strategies for cell therapies? They could overcome some of the cost and complexities associated with lentiviral vectors, but would perhaps be more limited by GMP plasmid production. Curious on your take there.
Are you asking about direct plasmid transfection/electroporation of transposases, or something like VLPs or exosomes? A few groups are working on using transposases to make cell therapies. Technologies like PASTE might be useful but are early-stage.
Here are some thoughts from our engineering teams:
There is probably a place in the market for lots of different methods, both non-viral (e.g. LNPs) and non-carrier (such as electroporation). The ideal method depends on the specific application (size of patient population, cell type being engineered, therapeutic dose, etc).
Benefits of non-viral methods include:
- Higher design flexibility; less limited by payload size.
- Maybe safer; avoids random integration typical of viral delivery.
- CRISPR-mediated integration is in theory safer, but still requires consideration of off-target insertion and may not achieve sufficient expression.
- CRISPR-HDR has some risk because of p53 pathway activation and other chromosomal integrity issues
- Some transposases may be biased toward intergenic regions, which would theoretically be safer than fully random/unbiased integration.
- (Sometimes) Lack of immunogenicity. But not all non-viral methods (e.g. VLPs) will lack immunogenicity.
Some typical drawbacks of non-viral delivery are:
- Cell loss/low viability (especially true for electroporation)
- Scalability
- Anything transfection-based will be difficult to scale.
Thank you for these thoughts! That all makes a lot of sense, and I certainly agree there is likely a market for both for many of the reasons you mentioned. I’m looking forward to reading more from your team on this front.
excellent ...article
Excellent article! I wonder what your team thinks about non-viral engineering strategies for cell therapies? They could overcome some of the cost and complexities associated with lentiviral vectors, but would perhaps be more limited by GMP plasmid production. Curious on your take there.
Are you asking about direct plasmid transfection/electroporation of transposases, or something like VLPs or exosomes? A few groups are working on using transposases to make cell therapies. Technologies like PASTE might be useful but are early-stage.
I was thinking along the lines of transposases or CRISPR-mediated homology-directed repair with a DNA repair template.
Here are some thoughts from our engineering teams:
There is probably a place in the market for lots of different methods, both non-viral (e.g. LNPs) and non-carrier (such as electroporation). The ideal method depends on the specific application (size of patient population, cell type being engineered, therapeutic dose, etc).
Benefits of non-viral methods include:
- Higher design flexibility; less limited by payload size.
- Maybe safer; avoids random integration typical of viral delivery.
- CRISPR-mediated integration is in theory safer, but still requires consideration of off-target insertion and may not achieve sufficient expression.
- CRISPR-HDR has some risk because of p53 pathway activation and other chromosomal integrity issues
- Some transposases may be biased toward intergenic regions, which would theoretically be safer than fully random/unbiased integration.
- (Sometimes) Lack of immunogenicity. But not all non-viral methods (e.g. VLPs) will lack immunogenicity.
Some typical drawbacks of non-viral delivery are:
- Cell loss/low viability (especially true for electroporation)
- Scalability
- Anything transfection-based will be difficult to scale.
Thank you for these thoughts! That all makes a lot of sense, and I certainly agree there is likely a market for both for many of the reasons you mentioned. I’m looking forward to reading more from your team on this front.