Type VII Collagen: The Anchoring Fibril Protein at Fault in Dystrophic Epidermolysis Bullosa

Chung, Hye Jin; Uitto, Jouni

doi:10.1016/j.det.2009.10.011

Cited by 153 publications

(120 citation statements)

References 41 publications

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“…After being secreted into the extracellular space, triple-helical COL7 molecules form antiparallel dimers. Subsequently, a large number of dimer molecules assemble laterally in register to form anchoring fibrils below the basement membrane zone (22). Given that secreted COL7 trimers are expressed from both keratinocytes and fibroblasts and could interfere with each other, it would be beneficial to gene-correct both keratinocytes and fibroblasts in a therapeutic strategy for DDEB.…”

Section: Discussionmentioning

confidence: 99%

“…It is known that three pro-COL7 polypeptides associate through their carboxyl-terminal ends within the intracellular space of keratinocytes and fibroblasts, and, subsequently, COL7 trimer is secreted into the extracellular space (22). To determine whether the gene-edited COL7 protein can be secreted appropriately, we generated a COL7 expression construct with the same mutation by sitedirected mutagenesis and assessed the ability of the recombinant COL7 to form trimers.…”

Section: Triple-helix Formation Analysis Of the Truncated Recombinantmentioning

confidence: 99%

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Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa

Shinkuma

Guo

Christiano

2016

Proc. Natl. Acad. Sci. U.S.A.

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Genome editing with engineered site-specific endonucleases involves nonhomologous end-joining, leading to reading frame disruption. The approach is applicable to dominant negative disorders, which can be treated simply by knocking out the mutant allele, while leaving the normal allele intact. We applied this strategy to dominant dystrophic epidermolysis bullosa (DDEB), which is caused by a dominant negative mutation in the COL7A1 gene encoding type VII collagen (COL7). We performed genome editing with TALENs and CRISPR/Cas9 targeting the mutation, c.8068_8084delinsGA. We then cotransfected Cas9 and guide RNA expression vectors expressed with GFP and DsRed, respectively, into induced pluripotent stem cells (iPSCs) generated from DDEB fibroblasts. After sorting, 90% of the iPSCs were edited, and we selected four gene-edited iPSC lines for further study. These iPSCs were differentiated into keratinocytes and fibroblasts secreting COL7. RT-PCR and Western blot analyses revealed gene-edited COL7 with frameshift mutations degraded at the protein level. In addition, we confirmed that the gene-edited truncated COL7 could neither associate with normal COL7 nor undergo triple helix formation. Our data establish the feasibility of mutation site-specific genome editing in dominant negative disorders.CRISPR/Cas | TALENs | epidermolysis bullosa | gene editing | dominant negative effect

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Section: Discussionmentioning

confidence: 99%

Section: Triple-helix Formation Analysis Of the Truncated Recombinantmentioning

confidence: 99%

Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa

Shinkuma

Guo

Christiano

2016

Proc. Natl. Acad. Sci. U.S.A.

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“…It is the most abundant type, being widely distributed in the body as structures classically referred to as collagen fibrils that form bones, dentin, tendons, capsules bodies, corneal and dermal blood vessels. These structures are known to play an important role in the morphogenesis and cell metabolism of new tissue, providing mechanical and biochemical properties (MYLLYHARJU and KIVIRIKKO, 2004;SÖDERHÄLL et al, 2007;CHUNG and UITTO, 2010;DABOOR et al, 2010;FERREIRA et al, 2012;MAKAREEVA and LEIKIN, 2014). TURKIEWIC et al (1991) isolated a serine proteinase from Euphausia superba with collagenolytic properties, which was able to hydrolyze collagens from Achilles tendon type I and release fibrils of calfskin collagen, under conditions that do not denature the substrates.…”

Section: Hydrolysis Of Collagen By the Collagenolytic Enzymementioning

confidence: 99%

Colagenase de pescada branca: extração, purificação parcial, caracterização e teste de especificidade ao colágeno para aplicação industrial

et al. 2017

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Fish processing residues are rich sources of biomolecules with industrial potential, such as enzymes with collagenolytic properties applied in the pharmaceutical, textile and leather sectors. Here, collagenolytic serine proteases were partially purified from the waste (viscera) of smooth weakfish Cynoscion leiarchus and characterized for the purpose of obtaining a value-added product from fisheries resources. The higher activity of the enzyme (72.5 U mL -1 ) was verified in optimal temperature and pH of 55°C and 8.0, respectively. The enzyme was stable in wide ranges of temperature (25-60°C) and pH (6.5 to 11.5). The ions Ca 2+ and Mg 2+ increased the protease activity, whilst Pb 2+, Al 3+ and Cu 2+ had an inhibitory effect, as observed in the presence of Benzamidine and TLCK (inhibitors of serine proteases). Hydrolysis was detected after 48 hours, when the enzyme and bovine collagen type I were incubated together. Thus, digestive viscera of C. leiarchus is suggested as an alternative source of enzymes capable of cleaving type I collagen, with similar biochemical properties to bacterial collagenases commonly employed in industrial processes, reducing costs, adding value to the fisheries product and minimizing the environmental impact caused by its waste. , com temperatura e pH ótimos de 55°C e 8.0, respectivamente. A enzima manteve-se estável em faixas amplas de temperatura (25-60 °C) e pH (6,(5)(6)(7)(8)(9)(10)(11)5 , Al 3+ e Cu 2+ inibiram essa atividade assim como os inibidores de serino-proteases (Benzamidina e TLCK). A hidrólise foi detectada após 48h de incubação com colágeno bovino tipo I. Assim, sugere-se vísceras digestivas de C. leiarchus como fonte alternativa para o fornecimento de enzimas com capacidade de clivar o colágeno do tipo I e com propriedades bioquímicas semelhantes às colagenases bacterianas já empregadas nas etapas de processamento industrial, como forma de redução de custo, agregação de valor ao produto pesqueiro e contribuindo para minimizar o impacto ambiental deste tipo de resíduo.

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“…Dystrophic EB could be inherited in both recessive and dominant form 1 . Several cases of DEB have been reported from India.…”

Section: Discussionmentioning

confidence: 99%

Case Report: Whole exome sequencing reveals a novel frameshift deletion mutation p.G2254fs in COL7A1 associated with autosomal recessive dystrophic epidermolysis bullosa

et al. 2016

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Dystrophic epidermolysis bullosa simplex (DEB) is a phenotypically diverse inherited skin fragility disorder. It is majorly manifested by appearance of epidermal bullae upon friction caused either by physical or environmental trauma. The phenotypic manifestations also include appearance of milia, scarring all over the body and nail dystrophy. DEB can be inherited in a recessive or dominant form and the recessive form of DEB (RDEB) is more severe. In the present study, we identify a novel p.G2254fs mutation in COL7A1 gene causing a sporadic case of RDEB by whole exome sequencing (WES). Apart from adding a novel frameshift Collagen VII mutation to the repertoire of known mutations reported in the disease, to the best of our knowledge, this is the first report of a genetically characterized case of DEB from India.

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Type VII Collagen: The Anchoring Fibril Protein at Fault in Dystrophic Epidermolysis Bullosa

Cited by 153 publications

References 41 publications

Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa

Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa

Colagenase de pescada branca: extração, purificação parcial, caracterização e teste de especificidade ao colágeno para aplicação industrial

Case Report: Whole exome sequencing reveals a novel frameshift deletion mutation p.G2254fs in COL7A1 associated with autosomal recessive dystrophic epidermolysis bullosa

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