How dangerous is the shRNA-expressing Lentivirus (possible contamination that could hurt young child)?

I supposed you are going to use PLKO.1 for your shRNA (3rd generation system).

The new package system consisted in 2 plasmid (+ yours shRNA): one encoding Rev and one encoding Gag and Pol which make the lentivirus production safer (Replication incompetent and always SIN). moreover once the infection has taken place it's impossible the production of new virions (if you pierce your skin just the cells in contact will be infected but you cannot transmit the virus).

You need to follow good practice rules: in our institute the lentivirus can be made just in a 2 class hood and VIRKON has to use in case of spillage and before and after your work; also double gloves, coat and tying your hair back are recommended. Lentivirus waste should be separated from other waste and autoclaved before disposing and the pipettes and tips should be washed with VIRKON before be autoclaved.

In case of small or medium spillage in the hood just wape down with 1% VIRKON; outside immediately spray the liquid with ethanol and put on top a tissue to avoid form any vapor. in case on spillage in the centrifuge close it and wait at least 30 min before re-opened and spray with VIRKON. Don't panic anyway in case of small spillage because the viruses quickly die when desiccated.

Good practice also suggests to have an allocated coat for this work and wash carefully the hands before leaving the facility.

Your baby will not be affected by lentivirus production at all.

PS I have more than 6 years experience with lentivirus and I have applied first for the licence in my institute. Also I'm one of the Lentivirus safety officer in my institution .

Lyra Chang

Dear Sanjay and Martina,

I apologize for my late reply!!

Thank you two for reminding me the importance of protection gears. I am especially grateful for your detailed reply, Martina! Now I am more confident in using Lentivirus for my research. I will be very careful during experiments and hopefully my baby will not be infected/contaminated by the viruses.

Thank you again and wish you two best of luck in your career & research!!

Best,

Lyra

Martina Anna Finetti

Lyra,

Pleasure!

I forgot to mention that you should avoid to use any sharp and glass (no syringes or glass pipettes) to avoid any cut and therefore possible virus contact.

This is very critical and it's one of the most common accident during Tc work

Lyra Chang

Thank you Martina! I will be very careful with the sharp items!

Do you know how long the lentivirus survive on any surface (without bleach)?

Thanks,

Lyra

Lyra Chang

Update: I have started my Lentivirus work and as an additional precaution, I changed clothes when I deal with the virus.

Also, share with you guys a very good guidelines for research with Lentivirus (see the following link).

Lyra Chang

The link for the guideline:

http://goo.gl/vxAcJH

Yu Wa

Dear Colleague,

The use of shRNA-expressing lentivirus in research laboratories is a common practice for gene silencing studies. These lentiviral vectors are engineered to minimize the risk to human health, incorporating several safety features to reduce their pathogenicity and replication capacity. However, it is essential to recognize that any exposure to lentiviral vectors, especially for vulnerable populations such as young children, should be avoided to prevent potential health risks.

Biosafety Considerations:Lentiviral vectors used in research are typically classified as Biosafety Level 2 (BSL-2) agents. This classification indicates that they pose moderate health risks and require specific containment measures to prevent accidental exposure.

Safety Features of Lentiviral Vectors:Modern lentiviral vectors are designed as self-inactivating (SIN) vectors, where crucial viral genes required for replication are deleted. This modification ensures that the virus can deliver genetic material into target cells without producing new infectious viral particles.

Potential Risks:Despite these safety measures, exposure to lentiviral vectors could potentially lead to the insertion of foreign genetic material into the genome of exposed individuals. While the risk of insertional mutagenesis (where the insertion of viral DNA disrupts normal gene function) is low, it cannot be entirely ruled out. Additionally, the immune response to viral proteins could pose health risks, especially for individuals with compromised immune systems or specific vulnerabilities, like young children.

Minimizing Exposure Risk: To minimize the risk of accidental exposure to lentiviral vectors:Follow all recommended biosafety protocols when handling lentiviral vectors, including the use of personal protective equipment (PPE), such as gloves and lab coats, and conducting work within a biosafety cabinet. Ensure that all waste materials are decontaminated and disposed of properly according to institutional and regulatory guidelines. Limit access to areas where lentiviral vectors are used and stored, especially for individuals not involved in the research, including children.

Emergency Procedures:Laboratories should have clear procedures in place for addressing accidental exposures, including immediate washing of the affected area, medical evaluation, and reporting the incident to appropriate safety officers.

In conclusion, while shRNA-expressing lentiviral vectors are engineered for safety and are widely used in research settings, it is crucial to adhere strictly to biosafety protocols to prevent accidental exposure. The risk to individuals, especially young children, can be minimized through proper laboratory practices, containment measures, and awareness of the potential hazards.

Best regards.

Reviewing the protocols listed here may offer further guidance in addressing this issue

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