shRNA/miRNA/gRNA rAAV Production Full Service, Pilot Scale
Features:
- Excellent gene delivery efficiency in most cell types including dividing and non-dividing or primary cells.
- Multiple serotypes (AAV-1, AAV-2, AAV-3, AAV-4, AAV-5, AAV-6, AAV-8, AAV-9, AAV-DJ/8 and AAV-DJ).
- Persistent transgene expression.
- Induce efficient gene silencing.
Large shRNA/miRNA/gRNA AAV cis vector inventory for constructing AAV cis plasmid with a specific promoter and reporter:
For Conventional shRNA/miRNA/TuD rAAV Production |
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Single-promoter shRNA/miRNA AAV cis vector
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Dual-promoter shRNA/miRNA rAAV cis vector
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Promoterless
CMV
CAG
H1
U6
Synapsin
UBC
EF1α
ALB(1.4)
ApoE/AAT1
CaMKII
ELA1
Enh358MCK
cTNT
GFAP
MBP
SST
TBG
αMHC
hRPE
mIP1
tMCK
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Promoterless
CMV
CAG
H1
U6
Synapsin
UBC
EF1α
ALB(1.4)
ApoE/AAT1
CaMKII
ELA1
Enh358MCK
cTNT
GFAP
MBP
SST
TBG
αMHC
hRPE
mIP1
tMCK
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CMV
CAG
H1
U6
Synapsin
UBC
EF1α
CaMKII
cTNT
GFAP
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eGFP
RFP
mRFP
mCherry
tdTOMATO
TurboGFP
eYFP
Venus
Luc
LacZ
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For CRISPR/Cas9 rAAV Production |
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Single-promoter CRISPR/Cas9 AAV cis vector
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Dual-promoter CRISPR/Cas9 AAV cis vector
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Promoterless
U6
H1
CMV
CAG
CBH
Synapsin
UBC
EF1α
ALB(1.4)
ApoE/AAT1
CaMKII
ELA1
Enh358MCK
cTNT
GFAP
MBP
SST
TBG
αMHC
hRPE
mIP1
tMCK
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Empty
CBH
U6
H1
sCMV
Syn
sEF1α
CK 0.4
cTNT
GFAP 0.6
sSyn
SST
TBG
αMHC
hRPE
mIP1
tMCK
CBH
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Cas9
hCas9
SpCas9
hSpCas9
SaCas9
AsCpf1
hAsCpf1
ftCas9
st1Cas9
cjCas9
cdCas9
ciCas9
ncCas9
st3Cas9
nmCas9
mgCas9
nsCas9
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Promoterless
sCMV
U6
H1
tRNA
Syn
sSyn
sEF1α
CaMKII 0.4
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CMV
sCMV
CBH
H1
U6
Syn
sSyn
sEF1α
CaMKII 0.4
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Cas9
hCas9
SpCas9
hSpCas9
SaCas9
AsCpf1
hAsCpf1
ftCas9
st1Cas9
cjCas9
cdCas9
ciCas9
ncCas9
st3Cas9
nmCas9
mgCas9
nsCas9
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Service Description:
1. Synthesize and clone shRNA into AAV cis vectors.
2. Pilot scale transfection of AAV·HT™ 293 cell into 10x150 mm plates.
3. Harvest rAAV followed by purification via advanced 2xCsCl ultra-centrifugation to obtain clinical trial grade of rAAV vector.
4. Desalting, filter sterilization, and AAV titration via qPCR.
rAAV Serotypes We Offer:
AAV-1, AAV-2, AAV-3, AAV-4, AAV-5, AAV-6, AAV-7, AAV-8, AAV-9, AAV-PHP.B, AAV-PHP.eB, AAV-DJ/8
and AAV-DJ.
Required Materials: NCBI accession #, target sequence, validated siRNA or validated shRNA sequence; miRNA accession #, pre-miRNA, mature miRNA or TuD (tough decoy) miRNA.
Turnaround Time: 3~4 weeks.
Deliverables: >0.1 mL super purified in vivo grade of rAAV vector at >1E+13 VG/mL*.
We offer discount for new customer, please request a quote with us today.
We also offer truncated custom shRNA/miRNA AAV service, please request a quote with us today.
Figure 1. A comparison of purity and infectivity of rAAV vectors from different sources showing super purified and super infectious (close to clinical trial grade) rAAV vector prepared via our advanced double CsCl ultra-centrifugation approach.
A. rAAV vectors (total 1E+9 VG per lane) from different sources were resolved on SDS-PAGE followed by silver staining. Lane 1: GMP manufactured rAAV vector from CHOP; Lane 2: rAAV prepared via our advanced 2xCsCl ultra-centrifugation approach; Lane 3: rAAV from Vector Core of BCM; Lane 4: rAAV from our competitor "V"; Lane 5: rAAV from our competitor "C"; Lane 6: Protein marker.
B. Super infectious rAAV vector prepared via advanced double CsCl ultra-centrifugation. Left panel: rAAV9-GFP (total 5E+9 VG) from our competitor "V" injected to mouse eye; Right panel: rAAV-9-GFP (total 5E+9 VG) purified via advanced 2xCsCl ultra-centrifugation injected to mouse eye.
Figure 2. A comparison of infectivity of rAAV vectors from different sources showing super purified rAAV prepared via advanced double CsCl ultra-centrifugation approach is super infectious.
rAAV1-GFP (total 2E+9 VG) from different sources were injected to mouse muscle tissue. The GFP fluorescence was visualized 3 weeks post injection. A. rAAV1-GFP from our competitor "V"; B. rAAV1-GFP from our pre-made rAAVs stock purified via advanced double CsCl ultra-centrifugation. C. Quantification data showed that our super purified rAAV (bar 2) is ~ 9 times more infectious than that (bar 1) prepared via conventional CsCl ultra-centrifugation.
* Final viral yield may depend on the nature of transgene.
