AAV Helper Free Packaging & Expression Systems

• Live helper virus 을 제거함으로써 안전하고 편리한 gene delivery system 입니다.
• AAV packaging을 위한 adenoviral genes 들은 pHelper plasmid (E2A, E4 and VA RNA)에 의해 제공되거나 293 packaging cells (E1)로부터 공급됩니다.
• 1-6까지의 native serotypes 및 AAV-DJ 와 AAV-DJ/8 군으로 공급됩니다.
• AAV-DJ 군은 8가지의 다른 native serotype (NAAV-2, AAV-4, AAV-5, AAV-8, AAV-9, avian AAV, bovine AAV, caprine AAV)을 조합하여 생성된 hybrid capsid를 제공함으로써, 다양한 tissue 와 cell types에서 native serotypes보다 높은 infection 효율을 보입니다.
• AAV-DJ/8군은 AAV-DJ의 heparin binding domain에 point mutation을 일으킨 제품으로, 이를 통해 생성된 recombinant AAV는 in vivo상에서 heart, brain tissue등에서 native serotypes 8 (AAV-8), 9 (AAV-9)와 유사하게 발현됩니다.
• Expression vector, GFP control vector, pHelper Vector 만 제공되는 Packaging System 및 전용 expression vector가 함께 제공되는 Expression System 이 있습니다.

제품의 구성

Components	Packaging System	Expression System
pAAV Vector	Yes	Yes
pHelper Vector	Yes	Yes
pAAV-GFP Control Vector	Yes	Yes
pAAV Expression Vector	-	Yes

Figure 1: Viral Gene Delivery using the AAV Helper Free System.

Figure 2: Production and Transduction of AAV2-GFP. Top Left: 293AAV cells prior to transfection (10X). Top Right: 293AAV cells 48 hours after transfection (10X). Bottom Left: GFP expression in 293AAV cells 48 hours after transfection (10X). Bottom Right: GFP expression in 293AD cells 48 hours after infection (20X).

Figure 3: Generation of AAV-DJ through Capsid DNA Family Shuffling.

Relative in vitro Infectivity of AAV Vectors

Cell Line	AAV-1	AAV-2	AAV-3	AAV-4	AAV-5	AAV-6	AAV-8	AAV-9	AAV-DJ	AAV-DJ/8
Huh-7	13	100	2.5	0	0.1	10	0.7	0	500	0.2
HEK293	25	100	2.5	0.1	0.1	5	0.7	0.1	500	0.3
HeLa	3	100	2	0.1	6.7	1	0.2	0.1	667	0.2
HepG2	3	100	16.7	0.3	1.7	5	0.3	ND	1250	0.5
Hep1A	20	100	0.2	1	0.1	1	0.2	0	400	0.1
911	17	100	11	0.2	0.1	17	0.1	ND	500	0
CHO	100	100	14	1.4	333	50	10	1	25000	5
COS	33	100	33	3.3	5	14	2	0.5	500	0.3
MeWo	10	100	20	0.3	6.7	10	1	0.2	2857	1
NIH3T3	10	100	2.9	2.9	0.3	10	0.3	ND	500	0.1
A549	14	100	20	ND	0.5	10	0.5	0.1	1000	0.1
HT1180	20	100	10	0.1	0.3	33	0.5	0.1	333	0.2
Monocytes	1111	100	ND	ND	125	1429	ND	ND	100	ND
Immature DC	2500	100	ND	ND	222	2857	ND	ND	200	ND
Mature DC	2222	100	ND	ND	333	3333	ND	ND	100	ND

• Adenoviral Expression
  • Premade Recombinant Adenoviruses
  • RAPAd® Adenoviral Expression Systems
• Lentiviral Expression
  • Premade Expression Lentiviral Particles
  • Pre-made mammalian stable cell lines
  • ViraSafe™ Lentiviral Packaging & Expression Systems