Blonde Centaur
about 1 year ago

Cas13 RNA design for knockdown of chicken cells

I have experiences in RNAi and Cas9 knockout experiments in mammalian cells, but am now facing a little challenge. I have primary chicken and duck endothelial cells that require knockdowns of genes - knockouts of these genes are impossible due to their importance for these cells. I have seen 1-2 papers looking at CMV/Ef1a...
Cas13CMVCRISPRGenetic EngineeringLentiCRISPRLentiCRISPR V2LentivirusMouse
Gold Gorgon
almost 4 years ago

CRISPR-CAS9 KO without reduction of mRNA (¿Continue with clons Isolation?)

I trying to generate a KO model in ha hTERT line of fibroblast using lentiCRISPR V2 with 3 gRNA. The gRNAs are desining to generate a delection between exons 1 and 3 of my gene. I already performance puromicine selection and try to validate expresion levels of mRNA for me gene in policlonal cell line but I don't get any cha...
CRISPRfibroblastGenetic EngineeringLentiCRISPRLentiCRISPR V2
Mauve Kappa
about 4 years ago

native lentiCRISPRv2 instead of lentiCRISPRv2 + sgRNA against GFP as a control: bad idea?

In a recent experiment I used lentiviral transduction of CRISPR to knock-out/down a gene of interest in a cell line. As a negative control I used the native lentiCRISPRv2 plasmid (with the 2kb filler) instead of the lentiCRISPRv2 plasmid cloned with a sgRNA against GFP for example. When I added the Puromycin to the cells, m...
CRISPRGenetic EngineeringLentiCRISPRLentivirusPuromycinsgRNA
Find your community. Ask questions. Science is better when we troubleshoot together.
Find your community. Ask questions. Science is better when we troubleshoot together.
Community Guidelines

Sci Find is a place for experimentation beyond the publication. We are a growing collaborative community of scientists and experimentalists from around the world who believe science is better when we work together.

  • Help us keep Sci Find a safe space for productive and collaborative conversations. Please be kind and respect your fellow community members. If you see abusive or alarming content, please report it to admin@scifind.net, and our team will take appropriate actions.
  • All information posted to this page is public and Open Access. Learn more about our commitment to Open Access by visiting our Docs.
  • We encourage our community to have collaborative troubleshooting discussions, and to share tips and tricks, experimental methods, resources, and opportunities.
  • You must be a member to create or engage with content. Sign Up to create your profile and join the community for free.
  • Follow a guild and create a post. Learn more about guilds here.
  • Post titles should be descriptive and include descriptive tags. Check out our tagging guidelines here.
Sign Up

Have a question?

Contact support@scifind.net or check out our support page.