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CpG-FREE DNA 6
pCpGfree-vitro - Selectable CpG-free Plasmids
Description
CpG-free Plasmid Backbone
pCpGfree-vitro is a family of expression vectors completely devoid of
CpG dinucleotides that are selectable in mammalian cells. Similarly to
the other pCpGfree plasmids, all the elements required for replication
and selection of the plasmids in bacteria, and gene expression in
mammalian cells have been modified to remove all CpG dinucleotides.
A Choice of Different Backbones
To better suit your needs, the pCpGfree-vitro plasmid comes with a
choice of selection: blasticidin, hygromycin or kanamycin/G418.
Each pCpGfree-vitro is available with a CpG-free allele of the LacZ gene
or a multiple cloning site (MCS). This MCS contains several commonly
used restriction sites, for convenient cloning of a CpG-free gene, such as
the genes provided in the pSELECT plasmid (see page 138) or any open
reading frame or cDNA.
Applications
Study the immunostimulatory effect of CpG motifs
pCpGfree-vitro plasmids represent innovative tools to study the effects
of CpG dinucleotides in a number of applications. DNA vaccination
exploits the immunostimulatory character of certain CpG motifs to prime
and boost the immune response. However, these immunostimulatory
CpG motifs are antagonized by CpG dinucleotides in certain distinct base
contexts, termed neutralizing CpG motifs. Both types of CpG motifs are
usually present in plasmidic DNA, and therefore may lead to an
unfavorable immune response. pCpGfree-vitro is the ideal tool to
overcome this problem, and may be used to study the effects of these
two types of CpG motifs by adding them in different configurations to
the pCpGfree-vitro backbone.
Study CpG methylation
CpG dinucleotides are key elements in a number of cellular functions
associated with chromatin. Several large multisubunit complexes,
consisting of methyl-CpG binding (MBD) proteins and histone
deacetylases, have been implicated in the regulation of chromatin
dynamics. These complexes are recruited to methylated CpG
dinucleotides by DNA methyl transferases (DNMTs) and induce
chromatin remodeling. However the specific roles of these complexes
are still to be explored. Due to the absence of CpG dinucleotides within
its backbone, pCpGfree-vitro is not the target of DNMTs and thus MBD
proteins. Therefore, it provides a useful model to study the other proteins
involved in these complexes, in particular the histone deacetylases. It can
also be used to analyze the effects of CpG methylation on the regulation
and duration of gene expression.
Contents and Storage
Each pCpGfree-vitro plasmid is provided as 20 μg of lyophilized DNA
with a paper disk of lyophilized E. coli GT115 strain. Products are shipped
at room temperature and should be stored at -20°C.
Selections Available in pCpGfree-vitro Plasmids
- Blasticidin
- Hygromycin
- Neomycin (G418)
PRODUCT QUANTITY CAT. CODE
pCpGfree-vitroBmcs 20 μg pcpgvtb-mcsg2
pCpGfree-vitroBLacZ 20 μg pcpgvtb-lz
pCpGfree-vitroHmcs 20 μg pcpgvth-mcsg2
pCpGfree-vitroHLacZ 20 μg pcpgvth-lz
pCpGfree-vitroNmcs 20 μg pcpgvtn-mcsg2
pCpGfree-vitroNLacZ 20 μg pcpgvtn-lz
Related Products
Important Licensing Information: These products are covered by a Limited Use License (See Terms and Conditions on page 164). By use of these products you
accept the terms and conditions of all applicable Limited Use Label Licenses.
136 www.invivogen.com/cpgfree-plasmid
Blasticidin, page 17 Hygromycin B, page 18
G418 Sulfate, page 17 LyoComp GT115, page 43
MCS
BsrGI
ScaI
BglII
ApaLI
Bsp120I
NcoI
NheI
MscI