pRetro-H1G vector, please submit your gene name, accession #, target and
loop sequences here.
conditions listed on the ordering page. Submitting an order form is construed as
you have carefully read and agree with such terms and conditions.
Purchase of the shRNA vector Construction service amounts to entering a service
agreement under which Cellogenetics is contractually obligated to:
1. synthesize the corresponding DNA oligoes for and generate the ordered shRNA
expression constructs.
2. confirm the cloned shRNA coding sequence by DNA sequencing.
3. make glycerol stocks of bacterial clones containing the ordered shRNA
expression constructs and prepare technical spec sheet.
4. ship the ordered shRNA expression constructs as glycerol stocks along with all
pertinent technical information within 15 business days from the date when the
order was placed.
designed to express double stranded small hairpin RNAs
from the human H1 RNA promoter. This vector is
constructed from the pMSCV retroviral vector with
backbone derived from the Murine Embryonic Stem Cell
Virus (MESV). A compounded puromycin resistance
internal ribosome entry site-green fluorescence protein
(PIG) cassette was inserted in place of the original simple
puromycin resistance gene behind the phosphoglycerate
kinase (PGK) promoter for antibiotic selection and green
fluorescence labeling in mammalian cells. Annealed
DNA oligoes for expression of small hairpin RNAs are to
be cloned behind the human H1 RNA promoter between
BamHI and Xho I sites. Upon transfection into a
permissive packaging cell line, such as Phoenix E cells,
pRetro-H1G expresses, either transiently or stably, viral
RNA transcripts to be packaged into infectious but
replication incompetent virus particles. The small
hairpin RNA encoded by pRetro-H1G can be delivered
either by standard transfection approach or by infecting
target cells with packaged virus particles. Either
approach achieves high level expression of small hairpin
RNAs in hematopoietic and embryonic stem cells, as well
as most other mitotically active cells.
All pRetro-H1G derived plasmids should be propagated in
suitable host strains such as DH5a, or HB101, under
selection of resistance to ampicillin (50 mg/ml). E.coli
replication origin: pUC; copy number: high.
