Enjoy Superior AAV Plasmid Expression
with GenScript’s New and Improved
AAV ITR Guarantee Technology

What are ITRs?
▪ Inverted terminal repeat (ITR): key element of an Adeno-associated virus (AAV) plasmid that is essential for theintegrity of AAV transfer plasmid.
▪ Each AAV plasmid has 2 ITR sequences (145 bases each) at each end flanking the transgene, forming a palindromic
hairpin structure that is high in GC content and unstable.
▪ This results in partial loss during plasmid propagation, with complete loss of ITR sequences abolishing AAVpackaging, greatly reducing AAV vector production to suboptimum levels.

ORFITRITR

GenScript’s Solution
Traditional solutions to prevent ITR loss are imperfect and does not promise conserved ITR integrity.
To overcome this issue, GenScript developed a Sanger-dependent ITR specific guarantee kit that can sequence
through the whole ITRs to determine its integrity with good signal.

Why AAV ITR Guarantee?
GenScript’s proprietary AAV ITR Guarantee technology
Regular sanger sequencing typically fails to sequence through ITR due to its high GC content, resulting intermination of sequencing reaction and subsequent incomplete sequencing. GenScript’s proprietary
AAV ITR Guarantee technology can easily read through ITR with clean peaks.

Traditional
Sanger-sequencing
AAV ITR Guarantee

Smal digestion does not always tell the truth
Restriction analysis using enzymes like Smal can be used as a rapid screen to detect ITR loss during
cloning experiments. However, traditional SmaI digestion does not always detect deletion in ITRs as smalldeletions within ITRs that don’t affect recognition sequences can’t be detected with these enzymes.
Highlighting a significant need for stringent ITR sequencing.
Mutations detected by AAV ITR Guarantee

Expected digestion band size

Band size on agarose gel

2.6 kb double band

2.6 kb double band

5.2 kb +1.1 kb

5.2 kb +1.1 kb

240 bp insertion

4.5 kb + 2.8 kb

4.5 kb + 2.8 kb

point mutation, 21 bp deletion

4.9 kb + 2.2 kb

4.9 kb + 2.2 kb

22 bp deletion

4.7 kb + 2.0 kb

4.7 kb + 2.0 kb

3 bp mutation, 22 bp deletion

Intact ITR

ITR with deletion

15 bp deletions

Agarose gel does not show deletion in ITR

GenScript’s proprietary strain out-performs commercial strains
Recombination-deficient commercial strains like NEBStbl, JC8111, SURE2, Stabl3, XL10-Gold are
frequently used for AAV construct amplifications, but are not always effective.
GenScript has developed a proprietary strain, GenITRv1, that has demonstrated improved ITR
integrity.
% of clones with intact ITR
2x7 plasmids, pick 8 clone each

120%
100%

89.29%

89.29%

80%
64.29%

60%
40%

44.64%

57.14%

44.64%

20%
0%
Stbl3

NEB Stbl
CloningGenlTR v1
Scale-up

Interested in the new service? Speak to a Ph.D. level specialist today!
Email us atGive us a call at

gene@genscript.com

1-877-436-7274 (Toll-Free) 1-732-885-9188

860 Centennial Ave., Piscataway, NJ 08854, USA | Toll-Free: 1-877-436-7274 | Telephone: 1-732-885-9188 | Fax: 1-732-210-0262
All Rights Reserved. All content described by GenScript is copyright of GenScript Corporation unless specifically identified otherwise. This includes all imagery, text andprogrammatic computer code.