Secreted Particle Information Transfer (SPIT) – A Cellular Platform for<i>In Vivo</i>Genetic Engineering

Charlesworth, Carsten T.; Homma, Shota; Suchy, Fabian; Wang, Sicong; Bhadhury, Joydeep; Amaya, Anais K.; Camarena, Joab; Zhang, Jinyu; Tan, Tze Kai; Igarishi, Kyomi; Nakauchi, Hiromitsu

doi:10.1101/2024.01.11.575257

Cited by 3 publications

(3 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Secreted Particle Information Transfer (SPIT) could also be exported for gene editing [47]. However, mRNA export may be more efficacious, as opposed to RNP secretion.…”

Section: Neuronal Gene Editingmentioning

confidence: 99%

Microglial Replacement and COURIER or SPIT for Neuronal Gene Editing

Renteln

2024

Preprint

View full text Add to dashboard Cite

Adeno-associated viral (AAV) vectors can be used for gene delivery. AAV.CAP-Mac was recently developed; it can cross the blood-brain barrier and transduce cells throughout the CNS. However, some brain regions were not transduced in adult rhesus monkeys. Additionally, AAV vectors can only package 5 kb of DNA. Also, AAV vectors may be genotoxic and cytotoxic, especially at high doses. Finally, AAV vectors are very expensive to produce at sufficiently high titers for treatment. Off-the-shelf cell-based delivery systems wherein the cells can infiltrate the target tissue and be hyper-motile would be ideal. Also, mRNA-based gene editing would be a transient treatment, and thus be much safer.

show abstract

“…Secreted Particle Information Transfer (SPIT) could also be exported for gene editing [47]. However, mRNA export may be more efficacious, as opposed to RNP secretion.…”

Section: Neuronal Gene Editingmentioning

confidence: 99%

Microglial Replacement and COURIER or SPIT for Neuronal Gene Editing

Renteln

2024

Preprint

View full text Add to dashboard Cite

show abstract

“…The edited TRMs could secrete proteins, RNA [104], and ribonucleoproteins [105] to surrounding cells.…”

Section: Periodic Whole-body Lipofuscin Removalmentioning

confidence: 99%

Towards periodic whole-body lipofuscin removal

Renteln

2024

Preprint

View full text Add to dashboard Cite

Lipofuscin is indigestible garbage that accumulates in the autophagic vesicles and cytosol of post-mitotic cells with age. Drs. Brunk and Terman postulated that lipofuscin accumulation is the main or at least a major driving factor in aging. They even posited that the evolution of memory is the reason why we get lipofuscin at all, as stable synaptic connections must be maintained over time, meaning that the somas of neurons must also remain in the same locale. In other words, they cannot dilute out their garbage over time through cell division. Mechanistically, their position certainly makes sense given that rendering a large percentage of a post-mitotic cell’s lysosomes useless must almost certainly negatively affect that cell and the surrounding microenvironment. Here, I explore the possibility that the accumulation of lipofuscin may to some extent exacerbate every other kind of age-related damage. I do not think that lipofuscin removal will reverse/prevent all forms of aging, just the major component facing us currently. In this piece, I will review what is known about lipofuscin accumulation from evolutionary and mechanistic standpoints and discuss a method of removing it from single cells in vitro.

show abstract

“…The viral protease matures HIV-1 virions and may also liberate Cas9 RNPs from Gag proteins but is unnecessary in murine leukemia virus-based particles packaging base editors. 32 Reverse transcriptase and integrase assemble with the capsid core to form the pre-integration complex for transgene integration. Removing either the viral protease (RT + INT) independently or the entire Pol polypeptide did not significantly decrease the activity of the EDVs, which shows that both Cas9 and Gag-Cas9 are functional (Fig.…”

Section: Removing Capsid Core-related Components Created Functional M...mentioning

confidence: 99%

Mechanism-guided engineering of a minimal biological particle for genome editing

Ngo,

Peukes,

Baldwin

et al. 2024

Preprint

View full text Add to dashboard Cite

The widespread application of genome editing to treat or even cure disease requires the delivery of genome editors into the nucleus of target cells. Enveloped Delivery Vehicles (EDVs) are engineered virally-derived particles capable of packaging and delivering CRISPR-Cas9 ribonucleoproteins (RNPs). However, the presence of lentiviral genome encapsulation and replication components in EDVs has obscured the underlying delivery mechanism and precluded particle optimization. Here we show that Cas9 RNP nuclear delivery is independent of the native lentiviral capsid structure. Instead, EDV-mediated genome editing activity corresponds directly to the number of nuclear localization sequences on the Cas9 enzyme. EDV structural analysis using cryo-electron tomography and small molecule inhibitors guided the removal of ~80% of viral residues, creating a minimal EDV (miniEDV) that retains full RNP delivery capability. MiniEDVs are 25% smaller yet package equivalent amounts of Cas9 RNPs relative to the original EDVs, and demonstrated increased editing in cell lines and therapeutically-relevant primary human T cells. These results show that virally-derived particles can be streamlined to create efficacious genome editing delivery vehicles that could simplify production and manufacturing.

show abstract

Secreted Particle Information Transfer (SPIT) – A Cellular Platform forIn VivoGenetic Engineering

Cited by 3 publications

References 25 publications

Microglial Replacement and COURIER or SPIT for Neuronal Gene Editing

Microglial Replacement and COURIER or SPIT for Neuronal Gene Editing

Towards periodic whole-body lipofuscin removal

Mechanism-guided engineering of a minimal biological particle for genome editing

Contact Info

Product

Resources

About