Yes, a Kozak sequence is essential for efficient translation and transgene expression in lentiviruses: In addition to the Kozak sequence, a woodchuck post-transcriptional regulatory element (WPRE) can also be added […]
Monthly Archives: October 2024
Primers and probes used for rAAV quantitative analysis
Applicability Target Sequences UniversalAll rAAV samples ITR2 primers: 5′-ggaacccctagtgatggagtt-3′ and5′-cggcctcagtgagcga-3′;probe: R-5′-cactccctctctgcgcgctcg-3′-Q. Semi-universalGroup of rAAV samples CMV promoter primers: 5′-ttcctacttggcagtacatctacg-3′ and5′-gtcaatggggtggagacttgg-3′;probe: R-5′-tgagtcaaaccgctatccacgccca-3′-Q;and other sequences. CAG promoter primers: 5′-ctgaccgcgttaatcccaca-3′ and5′-acaagccgtgattaaaccaaga-3′. CBA promoter […]
Quantification for residual host cell DNA contamination in the rAAV prep via qPCR or ddPCR analysis targeting highly repetitive genome sequences
DNA Impurity Target Sequences Plasmid KanR primers: 5′-gggcgcccggttctttttgtc-3′ and5′-gccagtcccttcccgcttcagtg-3′. AmpR primers: 5′-cgcgccacatagcagaactt-3′ and5′-cgccccgaagaacgttt-3′;probe: R-5′-aaaagtgctcatcattg-3′-Q;and other sequences. Adenovirus E2A primers: 5′-ttgctgaaacccaccatttg-3′ and5′-tcgtggacagcgaggaaga-3′;probe: R-5′-cgccacatcttctct-3′-Q. Adenovirus E4 primers: 5′-tcggcgcactccgtaca-3′ and5′-cgcgggtctctgtctcaaaa-3′;probe: R-5′-tagggatcgcctacctc-3′-Q. HSV UL23 […]
How to protect RNA from degradation?
To protect RNA from degradation, you can: RNA is prone to enzymatic degradation and hydrolysis. Even at low temperatures, RNA can still be reactive, and some ribozymes can be enhanced by […]
What is the main reason for RNA being unstable?
Unlike DNA, RNA is usually single-stranded. Additionally, RNA contains ribose sugars rather than deoxyribose sugars, which makes RNA more unstable and more prone to degradation. RNA is generally stable at –80° […]
Why Are Transgenes Cloned into Lentiviral Vectors Without a Poly(A) Signal?
In lentiviral gene delivery systems, transgene expression cassettes are commonly cloned without a polyadenylation signal to prevent disruption of full-length lentiviral genomes on mRNA maturation in producer cells. Since lentivirus is […]