Untitled

Lavado, Alfonso; Matheu, Ander; Serrano, Manuel; Montoliu, Lluı́s

doi:10.1186/1471-2121-6-18

Cited by 13 publications

(10 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These analyses need to be addressed in homozygous mice. Similarly, the possible alteration of chromatin marks around the Tyr locus will have to be addressed in immortalized melanocytes, following experimental approaches described in previous studies ( 68 ).…”

Section: Discussionmentioning

confidence: 99%

Functional validation of mouse tyrosinase non-coding regulatory DNA elements by CRISPR–Cas9-mediated mutagenesis

Seruggia

Fernández

Cantero

et al. 2015

Nucleic Acids Research

View full text Add to dashboard Cite

Newly developed genome-editing tools, such as the clustered regularly interspaced short palindromic repeat (CRISPR)–Cas9 system, allow simple and rapid genetic modification in most model organisms and human cell lines. Here, we report the production and analysis of mice carrying the inactivation via deletion of a genomic insulator, a key non-coding regulatory DNA element found 5′ upstream of the mouse tyrosinase (Tyr) gene. Targeting sequences flanking this boundary in mouse fertilized eggs resulted in the efficient deletion or inversion of large intervening DNA fragments delineated by the RNA guides. The resulting genome-edited mice showed a dramatic decrease in Tyr gene expression as inferred from the evident decrease of coat pigmentation, thus supporting the functionality of this boundary sequence in vivo, at the endogenous locus. Several potential off-targets bearing sequence similarity with each of the two RNA guides used were analyzed and found to be largely intact. This study reports how non-coding DNA elements, even if located in repeat-rich genomic sequences, can be efficiently and functionally evaluated in vivo and, furthermore, it illustrates how the regulatory elements described by the ENCODE and EPIGENOME projects, in the mouse and human genomes, can be systematically validated.

show abstract

Section: Discussionmentioning

confidence: 99%

Functional validation of mouse tyrosinase non-coding regulatory DNA elements by CRISPR–Cas9-mediated mutagenesis

Seruggia

Fernández

Cantero

et al. 2015

Nucleic Acids Research

View full text Add to dashboard Cite

show abstract

“…Cultures of immortal FMN1-deficient melanocytes (melan-f) were derived essentially 63 . In brief, mice carrying a previously generated FMN1 loss of function allele (FMN1pro) were crossed with Ink4a-Arf mutant mice, in order to generate pups homozygous for the FMN1pro mutant allele and heterozygous for Ink4a-Arf mutant allele.…”

Section: Methodsmentioning

confidence: 99%

Rab27a co-ordinates actin-dependent transport by controlling organelle-associated motors and track assembly proteins

et al. 2020

View full text Add to dashboard Cite

Cell biologists generally consider that microtubules and actin play complementary roles in long-and short-distance transport in animal cells. On the contrary, using melanosomes of melanocytes as a model, we recently discovered that the motor protein myosin-Va works with dynamic actin tracks to drive long-range organelle dispersion in opposition to microtubules. This suggests that in animals, as in yeast and plants, myosin/actin can drive longrange transport. Here, we show that the SPIRE-type actin nucleators (predominantly SPIRE1) are Rab27a effectors that cooperate with formin-1 to generate actin tracks required for myosin-Va-dependent transport in melanocytes. Thus, in addition to melanophilin/myosin-Va, Rab27a can recruit SPIREs to melanosomes, thereby integrating motor and track assembly activity at the organelle membrane. Based on this, we suggest a model in which organelles and force generators (motors and track assemblers) are linked, forming an organelle-based, cell-wide network that allows their collective activity to rapidly disperse the population of organelles long-distance throughout the cytoplasm.

show abstract

“…(melan-f) were derived essentially as described previously 59 . In brief mice carrying a previously generated FMN1 loss of function allele (FMN1 pro ) were crossed with Ink4a-Arf mutant mice in order to generate pups homozygous for the FMN1 pro mutant allele and heterozygous for Ink4a-Arf mutant allele.…”

Section: Derivation and Maintenance Of Cultured Cells Cultures Of Immentioning

confidence: 99%

“…In brief mice carrying a previously generated FMN1 loss of function allele (FMN1 pro ) were crossed with Ink4a-Arf mutant mice in order to generate pups homozygous for the FMN1 pro mutant allele and heterozygous for Ink4a-Arf mutant allele. Genotyping of the embryos was as previously described 37,59 . Melanocytes were then derived from the dorsal skin of mutant embryos as previously described 60 .…”

Section: Derivation and Maintenance Of Cultured Cells Cultures Of Immentioning

confidence: 99%

Rab27a co-ordinates actin-dependent transport by controlling organelle-associated motors and track assembly proteins

Alzahofi

Robinson

Welz

et al. 2018

Preprint

View full text Add to dashboard Cite

This 'highways and local roads' model suggests that MTs are tracks for long-range transport (highways) between the cell centre and periphery, driven by kinesin and dynein motors. Meanwhile AFs (local roads) and myosin motors work down-stream picking up cargo at the periphery and transporting it for the 'last m' to its final destination. This model makes intuitive sense as MTs in animal cells in culture typically form a polarised radial network of tracks spanning >10 m from the centrally located centrosome to the periphery and appear ideally distributed for long-distance transport. Meanwhile, with some exceptions in which AFs form uniformly polarised arrays, e.g. lamellipodia, filopodia and dendritic spines, AF architecture appears much more complex. In many fixed cells AF appear to comprise populations of short (1-2 m length), with random or anti-parallel filament polarity, and not an obvious system of tracks for directed transport 5,6 . This view is exemplified by the co-operative capture (CC) model of melanosome transport in melanocytes 7,8 . Skin melanocytes make pigmented melanosomes and then distribute them, via dendrites, to adjacent keratinocytes, thus providing pigmentation and photo-protection (reviewed in 9 ). The CC model proposes that transport of melanosomes into dendrites occurs by sequential longdistance transport from the cell body into dendrites along MTs (propelled by kinesin/dynein motors), followed by AF/myosin-Va dependent tethering in the dendrites. Consistent with this, in myosin-Va-null cells melanosomes move bi-directionally along MTs into dendrites, but do not accumulate therein, and instead cluster in the cell body 7,10 . This defect results in partial albinism in mammals due to uneven pigment transfer from melanocytes to keratinocytes (e.g. dilute mutant mouse and human Griscelli syndrome (GS) type I patients; Figure 1A) 11,12 . Subsequent studies revealed similar defects in mutant mice (and human GS types II and III patients) lacking the small

show abstract

Untitled

Cited by 13 publications

References 45 publications

Functional validation of mouse tyrosinase non-coding regulatory DNA elements by CRISPR–Cas9-mediated mutagenesis

Functional validation of mouse tyrosinase non-coding regulatory DNA elements by CRISPR–Cas9-mediated mutagenesis

Rab27a co-ordinates actin-dependent transport by controlling organelle-associated motors and track assembly proteins

Rab27a co-ordinates actin-dependent transport by controlling organelle-associated motors and track assembly proteins

Contact Info

Product

Resources

About