Enhancing sgRNA Expression in Mammalian Cells
Hi,
I'm working with sgRNA expression in mammalian cells using the U6 promoter. Are there stronger promoters or modifications to increase sgRNA or CRISPR array expression?
Thanks,
Anan
dCasRx/13 and splicing
dCasRx/13 and splicing
I am reaching out to inquire if anyone in the community has experience using dCasRx to target exons or introns, specifically in the context of observing changes in splicing patterns. If you have worked with dCasRx in this manner, I would be grateful if you could share your insights or any relevant resources.
Thank you in ad...
CRISPR ARRAY - Cas13
CRISPR ARRAY - Cas13
Hello!
Has anyone successfully performed Gibson assembly for six Cas13 guides? If so, did you incorporate the direct repeat sequence within your ordered guides?
I know how to do it to Cas9 sgRNAs but a little bit confusing for Cas13 because of the repetitive DR.
Any help will be highly appreciated.
Best,
Anan
Cas13 RNA design for knockdown of chicken cells
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...
Tool for crRNA design for cas13
Tool for crRNA design for cas13
Do you have any recommendations in terms of tools for guide RNA design for cas13? And does the crRNA of active cas13 works for deactivated cas13d.
New Research Team
New Research Team
Good afternoon, I am a researcher from Brazil and I’m initiating a study with crispr cas13 focusing on diagnosis. I wanted to ask if you have any advice that can help a beginner. I’m open to suggestions, in a matter of sellers or distributors that you could recommend, like the best place to get this plasmid, the best reagen...
How long the DR sequence should be for Lwa Cas13a crRNA?
How long the DR sequence should be for Lwa Cas13a crRNA?
ne, I want to design a RNA detect system, and for Lwa Cas13a crRNA design, I suppose I need a sequence with DR sequence + crRNA ( from 5 ends to 3 ends). From the literuatures, I found some of the design employed 36 nt sequence as the DR sequence, and some employed the DR sequence with 16 nt, if there a principle for DR se...
RNA targeting crRNA design at 5'UTR
RNA targeting crRNA design at 5'UTR
everyone!
Recently I designed four PspCas13b crRNAs at 5'UTR of KRAS, and made the KRAS crRNA pubulicated in Zhangfeng's paper as a positive control. When perform qPCR to detect the expression level of gene, only the positive control worked but four crRNAs. So I want to know whether the design principle of crRNA shoul...
Cas13 (C2C2) gRNA design tool
Cas13 (C2C2) gRNA design tool
I am interested in targeting mRNA with Cas 13. Is there an online tool to design gRNA for Cas13?
Cas13 system
Cas13 system
I was wondering if anyone has successfully used the cas13 (LwaCas13a, PspCas13b, or RfxCas13d) system in mammalian cells for gene know down. What are your experiences with these systems?
let me know.