First lentivirus is a subclass of retovirus. The reason is the nuclear import. The retrovirus such as MLV cannot transduce/infect non dividing cells because they cannot cross the nuclear membrane. Lentivirus can do that, their genome can go into the nucleus through nuclear pores.

Gene therapy replaces a faulty gene or adds a new gene in an attempt to cure disease or improve your body’s ability to fight disease. Gene therapy holds promise for treating a wide range of diseases, such as cancer, cystic fibrosis, heart disease, diabetes, hemophilia and AIDS.

Lentiviruses are RNA viruses that belong to the family Retroviridae. They contain the reverse transcriptase enzyme that converts RNA into DNA before becoming integrated into the genome of the host.

Most lentiviral vectors (LVVs) are derived from the human immunodeficiency virus (HIV) type 1 and retain the ability to integrate into the genome of infected cells. However, this powerful tool also carries the potential to cause oncogenic, infectious, and other transformative changes to infected cells.

Absolutely it is possible. I would recommend using a virus that is VSV-G pseudotyped (most commercially available ones are) as this will infect both mouse and rat (and also human if you want) cells with high efficiency.

Lentivirus is regarded as a biosafety level 2 material and safe to use due to its modified features (deletion of a number of accessory virulence genes , minimal genome of the viral particles, non-replicating and self-inactivation features), making it incapable of producing virus once infected into the host cell.

Lentivirus is a member of the retrovirus family, which also includes HIV-1. They are 80–100 nm in diameter and contain two copies of single-stranded RNA. Following entry into the cell, the RNA is reverse transcribed into linear double-stranded DNA in the cytoplasm.

c. Transduction

1. Thaw the lentivirus on ice. Mix 8 µl Polybrene (1 mg/ml aliquot) with 957 µl culture.
2. The next day, exchange Lentivirus/Polybrene mixture by fresh culture medium. Incubate cells at standard cell culture conditions.
3. concentrations range from 0.1-10 μg/ml. Replace the culture medium 48-72 hours.

Lentiviral vectors in gene therapy is a method by which genes can be inserted, modified, or deleted in organisms using lentivirus. Lentivirus are a family of viruses that are responsible for notable diseases like AIDS, which infect by inserting DNA into their host cells’ genome.

Lentiviruses (a genus of retrovirus) express reverse transcriptase, which converts the viral RNA to double stranded DNA, and integrase, which inserts this viral DNA into the host DNA. Once the viral DNA is integrated into the host DNA, it divides along with host cell and none are the wiser.

Add 4 µg/ml Polybrene and replace medium from target cells with the virus.

Depending on the construct used, transiently expressed transgene can generally be detected for 1 to 7 days, but transiently transfected cells are typically harvested 24 to 96 hours post-transfection. Analysis of gene products may require isolation of RNA or protein for enzymatic activity assays or immunoassays.

How do doctors and researchers decide whether a disease is a good candidate for gene therapy? They ask a couple of questions such as is the disease caused by gene mutation, Easier to cure the less genes that are mutated, can it be transferred by a vector, and what does the disorder do and will fixing the gene fix it.

2. What are the advantages and disadvantages of using viral vectors for gene therapy? –> Some disadvantages of using viral vectors are that the new gene may be inserted in the wrong DNA location. An advantage of using viral vectors is that some viruses a=can transfer good genes into the human cell.

Benefits and advantages of using the Adeno-Associated Virus as the vector in gene therapy trials include integration into host genome, no viral genes, able to transduce cells not actively dividing, wide range of host cells, and they are non-inflammatory and non-pathogenic.

SERCA2A. The knock-out mouse we received has a gene deletion for SERCA2a. If the SERCA2A knock-out mouse shows the same phenotype as the mouse with induced heart failure, we provided strong evidence that the SERCA2A gene is indeed involved in the development of heart failure.