Wei Li

10 Questions 7 Answers 0 Followers

Questions related from Wei Li

If I put 5'-CMV-cDNA-polyA-3' on 5' ITR-3'PolyA-cDNA-CMV3'-3'TR, can cDNA be expressed in rAAV vector?

I use homologous recombination cloning to assemble CMV-cDNA-PolyA (3.9 bp) to AAV transfer vector. After sequencing, I found the fragment is reversely inserted between 5'ITR and 3'ITR. We have...

15 May 2022 5,908 0 View

Is there a relation between CpG island in the expression persistence of AAV transgene and AAV titer?

For example, based on experience, the higher GC content in cDNA of AAV, relatively higher titer but lower expression persistence in vivo.

11 February 2022 4,102 3 View

Which codon optimization tool is good to enhance expression level and persistence of cDNA in rAAV?

ThermoFischers GeneArt Gene Synthesis NovoPro Labs Codon Optimization Tool (ExoOptimizer) Genewiz Codon Optimization ATUM DNA2.0 IDT Synthetic gene design made easy GenScript- GenSmart Codon...

11 February 2022 4,538 2 View

What are the possible drawbacks of using miRNA sponge to enhance the specificity of gene expression?

I am searching for miRNA repeats put between WPRE and 3'LTR of my lentivirus, in order to enhance the specificity of transgene expression in human neuron cells. But if clinically translation of...

27 December 2020 10,255 2 View

How can we sythesize 6 kb mRNA in vitro?

For in vivo method, can we engineer cells to produce exosomes carrying 6 kb mRNA ?? But in vitro method, maybe more straightforward.

20 November 2020 677 3 View

Lentivirus leave VSV-G and p26 protein on the surface of infected cells, then can immune cell (e.g. T cells) recognize viral protein to kill cell?

When we injected therapeutic lentivirus to a human cell in vivo, then lentivirus infects host cells, meanwhile leave VSV-G and p26 protein on the surface of infected cells, whether host immune...

14 March 2020 6,320 0 View

Do you have any recommended tools/companies that can design 5' UTR and 3' UTR?

I wanna add improved 5' UTR and 3' UTR to transgene in lentivirus to improve the stability/transcription/translation of transgene.

06 March 2020 2,709 0 View

What element(s) can replace WPRE but serve as almost same function as WPRE ?

I wanna put two WPRE in lentivirus vector. WPRE-transgene 1-promoter 1-promoter 2-transgene 2-WPRE. But I am worried two WPRE in lentivirus would reduce titre to great extent. So maybe I can...

06 March 2020 9,903 6 View

How to lyse cell for detecting L-levodopa and dopamine via HPLC?

We quite wanna know what the working concentration of sodium metabisulphite (Sigma) should be appropriate. sodium metabisulphite (Sigma) is antioxidant for protecting L-levodopa and dopamine...

12 February 2020 6,913 0 View

I am wondering whether hFEN1 or hExo1 can cleave off 5'-bases instead of 3'-flap??

To be specific, can I use hFEN1 or hExo1 (structure-specific enzyme) or other human enzymes to only remove a part of 5' ssDNA chain (blue line) in dsDNA genome of a human nucleus? Are there are...

03 November 2019 1,949 0 View