saporin

Ryan Y, Harrison A, Trivett H, Hartley C, David J, Clark GC, Hiscox JA (2022) RIPpore: A novel host-derived method for the identification of ricin intoxication through oxford nanopore direct RNA sequencing. Toxins (Basel) 14(7):470. doi: 10.3390/toxins14070470

Objective: The Depurination event could be detected using Oxford Nanopore Technologies (ONT) direct RNA sequencing, detecting a change in charge in the ricin loop.

Summary: Collectively, this work highlights the potential for ONT and direct RNA sequencing to detect and quantify depurination events caused by ribosome-inactivating proteins such as ricin.

Usage: Saporin was added as described by Rust et al., at 100 nM [22] for 24 h.

Related Products: Saporin (Cat. #PR-01)

See Also:

• Rust A et al. Two complementary approaches for intracellular delivery of exogenous enzymes. Sci Rep 5:12444, 2015.

Li Y, Ye Z, Yang H, Xu Q (2022) Tailoring combinatorial lipid nanoparticles for intracellular delivery of nucleic acids, proteins, and drugs. Acta Pharm Sin B 12(6):2624-2639. doi: 10.1016/j.apsb.2022.04.013

Objective: To highlight the recent progress in combinatorial lipid nanoparticles (LNPs) with novel structures and properties for the delivery of small- and macromolecular therapeutics.

Summary: The administration of protein/LNP negatively impacted reproduction in rats, including sperm production, estrous cyclicity and testicular and ovarian morphology, without causing any significant side effects. This non-surgical approach can be developed into a safe and convenient strategy for controlling the overproduction of pet and wildlife.

Usage: Intravenous administration of saporin loaded LNPs

Related Products: Saporin (Cat. #PR-01)

McDonough KE (2022) Maintenance mechanism of nociplastic pain in males. University of Texas Medical Branch Thesis.

Objective: The objective of this dissertation is to elucidate the sex-specific mechanisms underlying the transition to and maintenance of a nociplastic pain state using animal models.

Summary: This PhD dissertation investigates the mechanisms underlying the transition from acute to chronic nociplastic pain using murine models. The study finds that in males, spinal microglial activation driven by GABAergic disinhibition allows normally innocuous stimulation to induce a transition to nociplastic pain maintained by spinal microglia and proinflammatory cytokines.

Usage: Intrathecal injection of Saporin or Mac-1-SAP at 8.85 μM.

Related Products: Mac-1-SAP mouse/human (Cat. #IT-06), Saporin (Cat. #PR-01)

Ancheta LR, Shramm PA, Bouajram R, Higgins D, Lappi DA (2022) Saporin as a commercial reagent: its uses and unexpected impacts in the biological sciences-tools from the plant kingdom. Toxins (Basel) 14(3):184. doi: 10.3390/toxins14030184 PMID: 35324681

Summary: Saporin is a ribosome-inactivating protein that can cause inhibition of protein synthesis and causes cell death when delivered inside a cell. Development of commercial Saporin results in a technology termed ‘molecular surgery’, with Saporin as the scalpel. Its low toxicity (it has no efficient method of cell entry) and sturdy structure make Saporin a safe and simple molecule for many purposes. The most popular applications use experimental molecules that deliver Saporin via an add-on targeting molecule. These add-ons come in several forms: peptides, protein ligands, antibodies, even DNA fragments that mimic cell-binding ligands. Cells that do not express the targeted cell surface marker will not be affected. This review will highlight some newer efforts and discuss significant and unexpected impacts on science that molecular surgery has yielded over the last almost four decades. There are remarkable changes in fields such as the Neurosciences with models for Alzheimer’s Disease and epilepsy, and game-changing effects in the study of pain and itch. Many other uses are also discussed to record the wide-reaching impact of Saporin in research and drug development.

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Xu L, Füredi N, Lutter C, Geenen B, Pétervári E, Balaskó M, Dénes Á, Kovács KJ, Gaszner B, Kozicz T (2022) Leptin coordinates efferent sympathetic outflow to the white adipose tissue through the midbrain centrally-projecting Edinger-Westphal nucleus in male rats. Neuropharmacology 205:108898. doi: 10.1016/j.neuropharm.2021.108898

Objective: To show that leptin bound to neurons of the Edinger-Westphal nucleus (EWcp) stimulated STAT3 phosphorylation and increases urocortin 1 (Ucn1)-production in a time-dependent manner.

Summary: EWcp/LepRb/Ucn1 neurons respond to leptin signaling as well as control white adipose tissue size and fat metabolism without altering food intake.

Usage: Ablation of EWcp leptin receptor (LepRb) positive neurons with leptin-saporin. Either unconjugated saporin (53 ng in 80 nl MQ, or Leptin-SAP (90 ng in 80 nl MQ, was injected into the rat midbrain.

Related Products: Leptin-SAP (Cat. #IT-47), Saporin (Cat. #PR-01)

Mitdank H, Tröger M, Sonntag A, Shirazi NA, Woith E, Fuchs H, Kobelt D, Walther W, Weng A (2022) Suicide nanoplasmids coding for ribosome-inactivating proteins. Eur J Pharm Sci 170:106107. doi: 10.1016/j.ejps.2021.106107

Objective: To investigate the anti-proliferative activity of suicide-nanoplasmids.

Summary: In an in vivo neuroblastoma tumor model, treated mice showed a reduced tumor growth.

Usage: Design of a suicide nanoplasmid vector with saporin.

Related Products: Saporin (Cat. #PR-01)

Ujvári B, Pytel B, Márton Z, Bognár M, Kovács LÁ, Farkas J, Gaszner T, Berta G, Kecskés A, Kormos V, Farkas B, Füredi N, Gaszner B (2022) Neurodegeneration in the centrally-projecting Edinger-Westphal nucleus contributes to the non-motor symptoms of Parkinson’s disease in the rat. J Neuroinflammation 19(1):31. doi: 10.1186/s12974-022-02399-w

Objective: To investigate whether neuron loss and alpha-synuclein accumulation in the urocortin 1 containing (UCN1) cells of the centrally-projecting Edinger-Westphal (EWcp) nucleus is associated with anxiety and depression-like state in the rat.

Summary: Neurodegeneration of urocortinergic EWcp contributes to the mood-related non-motor symptoms in toxic models of Parkinson’s disease in the rat.

Usage: Leptin-SAP or unconjugated Saporin (0.08 μl) was injected into the EWcp area. This selective ablation of UCN1 neurons was used to validate the depression-like phenotype in rats. Behavioral, functional–morphological, biochemical and histopathological tools were used to test the motor coordination, mood status as well as morphological changes in the brain.

Related Products: Saporin (Cat. #PR-01), Leptin-SAP (Cat. #IT-47)

Hickerson BT, Daniels-Wells TR, Payes C, Clark LE, Candelaria PV, Bailey KW, Sefing EJ, Zink S, Ziegenbein J, Abraham J, Helguera G, Penichet ML, Gowen BB (2022) Host receptor-targeted therapeutic approach to counter pathogenic New World mammarenavirus infections. Nat Commun 13(1):558. doi: 10.1038/s41467-021-27949-3 PMID: 35091550

Objective: Demonstrate that a fusion protein of the antibody (ch128.1/IgG1) directed against the apical domain of human transferrin receptor 1 (hTfR1) and the Machupo virus (MACV) can inhibit infection of attenuated and pathogenic New World mammarenaviruses (NWMs).

Summary: NWMs cause life-threatening hemorrhagic fever (HF) and these viruses enter into cells via hTfR1. Use of ch128.1/IgG1 with other promising direct-acting small molecule antivirals or antibodies targeting the viral envelope glycoprotein would provide a complementary therapeutic strategy that would increase efficacy and reduce the emergence of drug resistance.

Usage: References MonoBiotin-ZAP reacted with avidinylated anti-hTfR (ch128.1Av) in a 1:1 molar ratio on ice for 30 minutes.

Related Products: MonoBiotin-ZAP (Cat. #BT-ZAP)

See Also:

• Daniels-Wells TR et al. Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1. Toxicol In Vitro 27(1):220-231, 2013.
• Daniels TR et al. Conjugation of an anti transferrin receptor IgG3-avidin fusion protein with biotinylated saporin results in significant enhancement of its cytotoxicity against malignant hematopoietic cells. Mol Cancer Ther 6:2995-3008, 2007.

Joshi BS, Ortiz D, Zuhorn IS (2021) Converting extracellular vesicles into nanomedicine: loading and unloading of cargo. Materials Today Nano 16:100148. doi: 10.1016/j.mtnano.2021.100148

Objective: To provide a comprehensive overview of (i) methods for the loading of EVs with therapeutic cargo, (ii) methods for EV surface functionalization to direct EVs to target cells, and (iii) methods to stimulate cargo release from EVs.

Summary: Development of methodologies to offload the natural cargo of EVs and substitute it for a cargo of interest, i.e., similar to the generation of empty viral capsids, may further aid in higher loading efficiency and enhanced therapeutic effects with improved safety.

Usage: Saporin, a small (30 kDa) ribosome-inactivating protein that induces cytotoxicity by inhibiting protein synthesis,was successfully loaded into EVs by electroporation.

Related Products: Saporin (Cat. #PR-01)

See Also:

• Nakase I Intracellular delivery methods using biofunctional peptide-modified extracellular vesicles. Extracell Vesicles Circ Nucleic Acids 1:44, 2020. American Society for Exosomes and Microvesicles 2020 Annual Meeting

Wu B, Zhao TV, Jin K, Hu Z, Abdel MP, Warrington KJ, Goronzy JJ, Weyand CM (2021) Mitochondrial aspartate regulates TNF biogenesis and autoimmune tissue inflammation. Nat Immunol 22(12):1551-1562. doi: 10.1038/s41590-021-01065-2

Objective: To identify a deficiency of mitochondrial aspartate production as a key abnormality in autoimmune T cells.

Summary: Shortage of mitochondrial aspartate disrupted the regeneration of the metabolic cofactor nicotinamide adenine dinucleotide (NAD), causing ADP-deribosylation of the endoplasmic reticulum (ER) sensor GRP78/BiP. Transfer of intact mitochondria into T cells as well as supplementation of exogenous aspartate rescued the mitochondria-instructed expansion of ER membranes and suppressed TNF release and rheumatoid tissue inflammation.

Usage: Immunofluorescence (Permeabilization by 0.5% Saporin)

Related Products: Saporin (Cat. #PR-01)