Regulation of the autophagic PI3KC3 complex by laforin/malin E3-ubiquitin ligase, two proteins involved in Lafora disease.

LaFora progressive myoclonus epilepsy is a rare neurodegenerative disorder characterized by the accumulation fatal dissolve abnormal glycogen deposits in the brain and peripheral tissues

. Mutations in at least two genes responsible for this disease: EPM2A, encoding laforin glucan phosphatase, and EPM2B, coding RING type E3-ubiquitin ligase master. Both laforin and malin form a functional complex where laforin recruit the substrate to ubiquitinated by the master. We and others have pointed out that, in cellular and animal models of the disease, there are disorders of autophagy that leads to the accumulation of dysfunctional mitochondria.

In addition, we determined that autophagic defect occurs in the early steps autophagosome formation. In this work, we provide evidence that the cellular models of disease there is a decrease in the number of phosphatidylinositol-3P. This may be due to defective regulation of autophagic PI3KC3 complex, in the absence of a functional laforin / master complex. In fact, we showed that laforin that / master complex physically interacts and co-localizes intracellularly to the core components of the complex PI3KC3 (Beclin1, Vps34 and Vps15), and that this interaction is specific and results in polyubiquitination this protein.

Additionally, laforin / malin complex is also capable polyubiquitinate ATG14L and UVRAG. Finally, we show that overexpression of laforin / complex increases the activity of master PI3KC3. These results demonstrate the new role of laforin / master complex in the activation of autophagy through regulation PI3KC3 complex and describes a defect in autophagy described in LaFora disease.
LaFora disease (LD) is a fatal epilepsy in children caused by a recessive mutation or gene either in EPM2A EPM2B.

A hallmark of LD is the accumulation of soluble polysaccharide intracellular deposits known as LaFora body (LBS) in the brain and other tissues. In the mouse model LD, genetic reduction of glycogen synthesis and rescue elimination LB formation neurological phenotype. Therefore, LBS has become a therapeutic target for ameliorating LD. Here, we show that human pancreatic α-amylase degradation of LBS. We amylase fused to an antibody fragment penetrates cells, and these antibody-enzyme fusion (VAL-0417) LBS degradation in vitro and dramatically reduce the burden of LB in vivo in Epm2a – / – mice.

 Regulation of the autophagic PI3KC3 complex by laforin/malin E3-ubiquitin ligase, two proteins involved in Lafora disease.
Regulation of the autophagic PI3KC3 complex by laforin/malin E3-ubiquitin ligase, two proteins involved in Lafora disease.

Delivery of the central nervous system and analysis of the biodistribution of the antibody-enzyme Fusion for Treatment of Diseases LaFora.

LaFora disease (LD) is a fatal epileptic teenager characterized by the accumulation of aberrant aggregates called LaFora body glucan (LBS). Protein-based therapeutic delivery to the central nervous system (CNS) for cleaning LBS remains a unique challenge in the field.

More recently, a humanized antigen binding fragment (hFab) derived from murine systemic lupus erythematosus DNA autoantibodies (3E10) has been shown to mediate cell penetration and is proposed as a carrier widely applicable to mediate cellular targeting and uptake. We report a study of the efficacy and CNS delivery of VAL-0417, the antibody-enzyme fusion consists of 3E10 hFab and human pancreatic α-amylase, in a mouse model of LD. Enzyme-linked immunosorbent assay was developed to detect VAL-0417 post-treatment as a measure of success of the post.

Frit Kit

FRIT-KIT 1each
EUR 124
Description: Kit to create frits in capillaries. Includes formamide, Kasil-1, Kasil-1624 and a cleaving tool.

Column Packing Kit

PACK-KIT 1pack
EUR 1035
Description: Column packing kit for pressure cells. Includes: HPREG regulator, TBNG10 tubing, CAP-75 capillary, and STRB5X2 stir bar.

PCR Mycoplasma Detection Kit

M034-Kit Kit
EUR 266

Cas9 Protein and T7 gRNA SmartNuclease Synthesis Kit

CAS400A-KIT 1 kit (10 rxn)
EUR 1110

CMV-hspCas9-T2A-Puro SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit

CASLV100PA-KIT 1 Kit
EUR 1132

CMV-hspCas9-EF1-GFP SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit

CASLV105PA-KIT 1 Kit
EUR 1132

MSCV-hspCas9-T2A-Puro SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit

CASLV120PA-KIT 1 Kit
EUR 1132

MSCV-hspCas9-EF1-GFP SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit

CASLV125PA-KIT 1 Kit
EUR 1132

Multiplex gRNA Kit + EF1-T7-hspCas9-H1-gRNA linearized SmartNuclease vector

CAS700A-KIT 10 rxn
EUR 1132

Multiplex gRNA Kit + CAG-T7-hspCas9-H1-gRNA linearized SmartNuclease vector

CAS720A-KIT 10 rxn
EUR 1132

Multiplex gRNA Kit + CMV-T7-hspCas9-H1-gRNA linearized SmartNuclease vector

CAS740A-KIT 10 rxn
EUR 1132

T7 gRNA SmartNuclease Synthesis Kit (includes CAS510A-1 & T7 IVT synthesis reagents)

CAS510A-KIT 1 Kit
EUR 805

Cas9 Nickase: CMV-hspCas9(D10A)-T2A-Puro SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit

CASLV200PA-KIT 1 Kit
EUR 1132

Cas9 Nickase: CMV-hspCas9(D10A)-EF1-GFP SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit

CASLV205PA-KIT 1 Kit
EUR 1132

Cas9 Nickase: MSCV-hspCas9(D10A)-T2A-Puro SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit

CASLV220PA-KIT 1 Kit
EUR 1132

Cas9 Nickase: MSCV-hspCas9(D10A)-EF1-GFP SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit

CASLV225PA-KIT 1 Kit
EUR 1132

Multiplex gRNA Kit + Cas9 Nickase: EF1-T7-hspCas9-nickase-H1-gRNA linearized SmartNickase vector

CAS750A-KIT 10 rxn
EUR 1132

Multiplex gRNA Kit + Cas9 Nickase: CAG-T7-hspCas9-nickase-H1-gRNA linearized SmartNickase vector

CAS770A-KIT 10 rxn
EUR 1132

Multiplex gRNA Kit + Cas9 Nickase: CMV-T7-hspCas9-nickase-H1-gRNA linearized SmartNickase vector

CAS790A-KIT 10 rxn
EUR 1132

Cas9 SmartNuclease Extra Ligation Kit [includes 5x ligation buffer (10 ul) and Fast ligase (2.5ul)]

CAS9LIG-KIT 1 Kit
EUR 153

PinPoint-FC 293T Platform Kit for Targeted Gene Insertion (includes PIN320A-1, PIN200A-1, PIN510A-1 & PIN600A-1)

PIN320A-KIT 1 Kit
EUR 4941

PinPoint-FC Murine iPSC Platform Kit for Targeted Gene Insertion (includes PIN340iPS-1, PIN200A-1, PIN510A-1 & PIN600A-1)

PIN340iPS-KIT 1 Kit
EUR 4941

AAVS1 Safe Harbor Targeting Vector 2.0 - All-Purpose Donor (AAVS1-SA-puro-MCS), Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector) and GE640PR-1 (Junction PCR Primer Mix to confirm AAVS1 integration site)

GE620A-KIT 1 kit
EUR 2132

AAVS1 Safe Harbor Targeting Vector 2.0 - GOI Knock-in Donor (AAVS1-SA-puro-EF1-MCS), Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector) and GE640PR-1 (Junction PCR Primer Mix to confirm AAVS1 integration site)

GE622A-KIT 1 kit
EUR 2132

AAVS1 Safe Harbor Targeting Vector 2.0 - Reporter Knock-in Donor (AAVS1-SA-puro-MCS-GFP), Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector) and GE640PR-1 (Junction PCR Primer Mix to confirm AAVS1 integration site)

GE624A-KIT 1 kit
EUR 2132

vWF Acty. Kit

ABP-ACT-KIT 12 x 8 microwells
EUR 428

vWF Ant. Kit

ABP-TOT-KIT 12 x 8 microwells
EUR 394

hspCas9 AAVS1 Safe Harbor Knock-in Donor (AAVS1-SA-puro-EF1-hspCas9)

CAS620A-KIT 1 kit
EUR 2152
Description: Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector), CAS640PR-1 (Junction PCR Primer Mix to confirm Cas9 integration site), and CAS9-PR-1 (PCR primers to confirm Cas9 expression)

PinPoint-FC System for Platform Cell Line Generation & Retargeting (includes PIN300A-1, FC200PA-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN300A-KIT 1 Kit
EUR 2798

PinPoint-HR System for Platform Cell Line Generation & Retargeting (includes PIN400A-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN400A-KIT 1 Kit
EUR 2798

PinPoint-HR System for Platform Cell Line Generation & Retargeting of AAVS1 Safe Harbor Locus (includes PIN410A-1, GE601A-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN410A-KIT 1 Kit
EUR 4335

PinPoint-HR System for Platform Cell Line Generation & Retargeting of AAVS1 Safe Harbor Locus (includes PIN410A-1, CAS601A-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN412A-KIT 1 Kit
EUR 4335

PrecisionX Multiplex gRNA Cloning Kit

CAS9-GRNA-KIT 10 rxn
EUR 445

mRNAExpress mRNA Synthesis kit (5 reactions)

MR-KIT-1 5 reactions
EUR 1152

ExoAb Antibody Kit (CD9, CD63, CD81, Hsp70 antibodies, rabbit anti-human) with goat anti-rabbit HRP secondary antibody

EXOAB-KIT-1 25 ul each
EUR 627

CLOuD9 Gene Expression Regulation Kit (includes 10 ug each of dCas9-PYL1 and dCas9-ABI1 lentivectors, and 100 ul of 0.5M Inducer Agent)

CASCL9-100A-KIT 1 Kit
EUR 1132

AXYGEN® AXYPET® PRO STARTER KIT: 4 PIPETTORS, PLEXI STAND, SAMPLE TIPS

AP-STR-KIT-P 1/pk
EUR 721
Description: Corning and Axygen Liquid Handling Equipment; Axypet Pipettors and Motopet Pipet Controller

AXYPET STARTER KIT 1 AP-20, AP-200 & AP-1000 WITH ADDITIONAL FREE RACKS OF AXYGEN PIPETTE TIPS

AP-STR-KIT-1 1/pk
EUR 355
Description: Corning and Axygen Liquid Handling Equipment; Axypet Pipettors and Motopet Pipet Controller

AXYPET STARTER KIT 2 AP-10, AP-100 & AP-1000 WITH ADDITIONAL FREE RACKS OF AXYGEN PIPETTE TIPS

AP-STR-KIT-2 1/pk
EUR 367
Description: Corning and Axygen Liquid Handling Equipment; Axypet Pipettors and Motopet Pipet Controller

Beta2-Microglobulin ELISA kit ELISA Kit

LF-EK60047 1×96T
EUR 679

Human Kit ELISA Kit

ELA-E0121h 96 Tests
EUR 824

LZM ELISA Kit| Mouse Lysozyme ELISA Kit

EF013748 96 Tests
EUR 689

MAU ELISA Kit| Mouse Microalbuminuria ELISA Kit

EF013752 96 Tests
EUR 689

MHB ELISA Kit| Mouse Methemoglobin ELISA Kit

EF013756 96 Tests
EUR 689

MT ELISA Kit| Mouse Melatonin ELISA Kit

EF013768 96 Tests
EUR 689

MTL ELISA Kit| Mouse Motilin ELISA Kit

EF013769 96 Tests
EUR 689

MYO ELISA Kit| Mouse Myoglobin ELISA Kit

EF013773 96 Tests
EUR 689

NGB ELISA Kit| Mouse Neuroglobin ELISA Kit

EF013783 96 Tests
EUR 689

Pg ELISA Kit| Mouse Progesterone ELISA Kit

EF013825 96 Tests
EUR 689

PHB ELISA Kit| Mouse Prohibitin ELISA Kit

EF013829 96 Tests
EUR 689

PI ELISA Kit| Mouse Proinsulin ELISA Kit

EF013830 96 Tests
EUR 689

PP ELISA Kit| Mouse Pepsin ELISA Kit

EF013842 96 Tests
EUR 689

PS ELISA Kit| Mouse Phosphatidylserine ELISA Kit

EF013849 96 Tests
EUR 689

PTD ELISA Kit| Mouse Pentosidine ELISA Kit

EF013851 96 Tests
EUR 689

PTN ELISA Kit| Mouse Pleiotrophin ELISA Kit

EF013856 96 Tests
EUR 689

REN ELISA Kit| Mouse Renin ELISA Kit

EF013865 96 Tests
EUR 689

SCT ELISA Kit| Mouse Secretin ELISA Kit

EF013878 96 Tests
EUR 689

SYP ELISA Kit| Mouse Synaptophysin ELISA Kit

EF013911 96 Tests
EUR 689

TG ELISA Kit| Mouse Thyroglobulin ELISA Kit

EF013933 96 Tests
EUR 689

TLN ELISA Kit| Mouse Talin ELISA Kit

EF013937 96 Tests
EUR 689

TM ELISA Kit| Mouse Thrombomodulin ELISA Kit

EF013939 96 Tests
EUR 689

TRF ELISA Kit| Mouse Transferrin ELISA Kit

EF013948 96 Tests
EUR 689

TRY ELISA Kit| Mouse Trypsin ELISA Kit

EF013951 96 Tests
EUR 689

TTR ELISA Kit| Mouse Transthyretin ELISA Kit

EF013954 96 Tests
EUR 689

Ub ELISA Kit| Mouse Ubiquitin ELISA Kit

EF013956 96 Tests
EUR 689

VF ELISA Kit| Mouse Visfatin ELISA Kit

EF013963 96 Tests
EUR 689

CCK ELISA Kit| Mouse Cholecystokinin ELISA Kit

EF013984 96 Tests
EUR 689

His ELISA Kit| Mouse Histamine ELISA Kit

EF013995 96 Tests
EUR 689

AMPKa ELISA Kit| Mouse pThr172 ELISA Kit

EF013997 96 Tests
EUR 689

PCT ELISA Kit| Mouse Procalcitonin ELISA Kit

EF014005 96 Tests
EUR 689

TE ELISA Kit| Mouse Telomerase ELISA Kit

EF014029 96 Tests
EUR 689

FUR ELISA Kit| Mouse Furin ELISA Kit

EF014041 96 Tests
EUR 689

Calprotectin ELISA Kit| Mouse Calprotectin ELISA Kit

EF014057 96 Tests
EUR 689

Hyp ELISA Kit| Mouse Hydroxyproline ELISA Kit

EF014063 96 Tests
EUR 689

FLG ELISA Kit| Mouse Filaggrin ELISA Kit

EF014071 96 Tests
EUR 689

Ahi1 ELISA Kit| Mouse Jouberin ELISA Kit

EF014105 96 Tests
EUR 689

Aaas ELISA Kit| Mouse Aladin ELISA Kit

EF014113 96 Tests
EUR 689

Adig ELISA Kit| Mouse Adipogenin ELISA Kit

EF014153 96 Tests
EUR 689

Afm ELISA Kit| Mouse Afamin ELISA Kit

EF014169 96 Tests
EUR 689

Ambn ELISA Kit| Mouse Ameloblastin ELISA Kit

EF014190 96 Tests
EUR 689

Atrn ELISA Kit| Mouse Attractin ELISA Kit

EF014265 96 Tests
EUR 689

Bsnd ELISA Kit| Mouse Barttin ELISA Kit

EF014276 96 Tests
EUR 689

Bfsp1 ELISA Kit| Mouse Filensin ELISA Kit

EF014298 96 Tests
EUR 689

Bfsp2 ELISA Kit| Mouse Phakinin ELISA Kit

EF014299 96 Tests
EUR 689

Bscl2 ELISA Kit| Mouse Seipin ELISA Kit

EF014300 96 Tests
EUR 689

Bgn ELISA Kit| Mouse Biglycan ELISA Kit

EF014309 96 Tests
EUR 689

Btd ELISA Kit| Mouse Biotinidase ELISA Kit

EF014312 96 Tests
EUR 689

Bche ELISA Kit| Mouse Cholinesterase ELISA Kit

EF014352 96 Tests
EUR 689

Ccin ELISA Kit| Mouse Calicin ELISA Kit

EF014375 96 Tests
EUR 689

Clgn ELISA Kit| Mouse Calmegin ELISA Kit

EF014376 96 Tests
EUR 689

Cdca8 ELISA Kit| Mouse Borealin ELISA Kit

EF014455 96 Tests
EUR 689

Cntln ELISA Kit| Mouse Centlein ELISA Kit

EF014458 96 Tests
EUR 689

Cer1 ELISA Kit| Mouse Cerberus ELISA Kit

EF014463 96 Tests
EUR 689

Chad ELISA Kit| Mouse Chondroadherin ELISA Kit

EF014487 96 Tests
EUR 689

Chodl ELISA Kit| Mouse Chondrolectin ELISA Kit

EF014490 96 Tests
EUR 689

Clspn ELISA Kit| Mouse Claspin ELISA Kit

EF014504 96 Tests
EUR 689

We showed robust and sensitive detection of the fusion protein in several types of tissue. By using this method, we measure biodistribution in a different method of delivery. We found that the intracerebroventricular administration provided strong CNS delivery when compared with intrathecal administration. These data define the essential steps in the translation pipeline VAL-0417 for the treatment of LD.