shRNA/miRNA/gRNA rAAV Production Full Service, Large Scale

Catalog #: SL100810
Unit Size: 1.0 mL at >10¹³VG/mL

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
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
CRISPR / Cas9 rAAV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. Design, synthesize and clone shRNA/miRNA into AAVcisvectors.
2. Large scale transfection ofAAV·HT™ 293 cells into 2xcell stack.
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 #, shRNA target sequence, validated siRNA or validated shRNA sequence; miRNA accession #, pre-miRNA or mature miRNA or TuD (tough decoy) miRNA.

Turnaround Time:3~4 weeks.

Deliverables:>1.0 mLsuperpurified in vivo grade rAAV vector at >1E+13 VG/mL*.

We offer discount for new customer, pleaserequest a quotewith us today.

We also offer truncated custom shRNA 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. Super infectious rAAV vector prepared via advanceddouble 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 2xCsClultra-centrifugationinjected 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 advanceddouble 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.