Houbaviy HB, Burley SK: Thermodynamic analysis of the interaction between YY1 and the AAV P5 promoter initiator element. Chem Biol 2001, 8 (2) : 179-187. PubMed Abstract Publisher Full TextHoubaviy H.B., Burley S.K. Thermodynamic analysis of the interaction between YY1 and the ...
(Figure 2A) Without helper virus, expression of Rep68/Rep78 would be restricted owing to Ying Yang 1 (YY1) repression of the P5 promoter, leading to inhibition of AAV genome replication and gene expression, and initiation of AAV chromosome integration (Figure 2B) [38]. AAV establishes ...
palindromic sequences that confer a DNA hairpin structure requiredin cisfor replication and encapsidation of the single-stranded viral genome6,7. The use of stable rep/cap cell lines, generally based on HeLa cells, has been explored extensively as a scalable packaging system for rAAV manufacture....
在哺乳动物细胞中的wtaav感染后,rep基因25(即rep78和rep52)分别从p5启动子和p19启动子表达,两种rep蛋白都在病毒基因组复制中起作用。reporf的剪接事件导致实际上4种rep蛋白(即rep78、rep68、rep52和rep40)的表达。然而,显示编码rep78和rep52蛋白的未剪接mrna在哺乳动物中足够用于aav载体生成。哺乳动物细胞中的...
2C, binding of Rep78 to the p5 promoter fragment was also enhanced by E1, and again, E1 alone did not bind p5. Download: Download high-res image (942KB) Download: Download full-size image Fig. 2. Enhanced binding of Rep78 to multiple AAV DNA substrates in the presence of E1. 32P...
The p5 and p19 promoters direct the synthesis of the viral regulatory proteins, Rep78 and -68 and Rep52 and -40, respectively. The p5 Rep proteins bind a linear 22-bp sequence, the Rep binding element (RBE), that is within both the terminal repeat (TR) and the p5 promoter. In the...
在哺乳动物细胞感染野生型(wt)AAV之后,Rep基因(即Rep78和Rep52)分别从P5启动子和P19启动子表达,这两个Rep蛋白均具有在病毒基因组中复制的功能。在RepORF中的剪接事件导致实际上四种蛋白质(即Rep78、Rep68、Rep52和Rep40)表达。然而,已经示出在哺乳动物细胞中编码Rep78和Rep52蛋白的未剪接的mRNA对于AAV载体产生...
The right side of a positive-sensed AAV genome encodes overlapping sequences of three capsid proteins, VP1, VP2 and VP3, which start from one promoter, designated p40. The molecular weights of these proteins are 87, 72 and 62 kiloDaltons, respectively. All three of them are translated from...
The AAV2 REP ORFs (Rep78, Rep68, Rep52, Rep40) were synthetized and codon optimized for human cell expression (Genscript, Piscataway, NJ) (sequences provided in supplementary information, Figure S4). Mutations were introduced in the internal AAV p19 promoter (TATTTAA to TACCTCA) for REP...
In one embodiment, an AAV P5 promoter sequence is employed. The selection of the AAV to provide any of these sequences does not limit the invention. In another preferred embodiment, the promoter for rep is an inducible promoter, such as are discussed above in connection with the transgene reg...