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和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 |
Single-promoter shRNA/miRNA AAV cis vector
 |
双-promoter shRNA/miRNA rAAV cis vector
 |
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 |
|
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 |
CMV
CAG
H1.
U6
Synapsin
UBC
EF1α
CaMKII
CTNT.
GFAP
|
egfp.
RFP
mRFP
mCherry
tdTOMATO
TurboGFP
eYFP
Venus
Luc.
LacZ
|
适用于CRISPR / CAS9 rav生产 |
Single-promoter CRISPR/Cas9 AAV cis vector
 |
双-promoter CRISPR/Cas9 AAV cis vector
 |
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 |
Empty
CBH
U6
H1.
SCMV
Syn
sEF1α
CK 0.4
CTNT.
GFAP 0.6
sSyn
SST.
TBG
αMHC
hRPE
mIP1
tMCK
CBH |
Cas9
hCas9
SpCas9
HSPCAS9.
SaCas9
AsCpf1
hAsCpf1
ftCas9
st1Cas9
CJCAS9.
cdCas9
ciCas9
ncCas9
st3Cas9
nmCas9
mgCas9
nsCas9
|
Promoterless
SCMV
U6
H1.
tRNA
Syn
sSyn
sEF1α
CaMKII 0.4 |
CMV
SCMV
CBH
H1.
U6
Syn
sSyn
sEF1α
CaMKII 0.4
|
Cas9
hCas9
SpCas9
HSPCAS9.
SaCas9
AsCpf1
hAsCpf1
ftCas9
st1Cas9
CJCAS9.
cdCas9
ciCas9
ncCas9
st3Cas9
nmCas9
mgCas9
nsCas9
|
Service Description:
1. Synthesize and clone shRNA into AAVcisvectors.
2. Pilotscale 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毫升超级纯化in vivo grade ofrAAV vector at >1E+13 VG/mL*.
We offer discount for new customer, pleaserequest a quotewith us today.
We also offer truncated custom shRNA/miRNA AAV service, pleaserequest a quotewith 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.通过先进准备的超级感染rav矢量双CSCL超离心. 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 2xCsClultra-centrifugation注射到小鼠眼睛。
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双CSCL超离心. 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.
