Genome Engineering using CRISPR/Cas Systems

Hello all, Did anyone try transfection of pCMV-AN-HA vector with an insert of gene of interest in

These genes may be essential for cell survival, have you considered that? If that's the case, you

Id choose ROSA26 murine locus, as many publications validate it as a safe harbour locus for transgene

Hi everyone. The CRISPR plasmid pX 333 from Zhang lab has been optimised for cloning of 2 single

Bumping this to the top - curious if anyone has any input on a best practice for these libraries that

Hi Everyone, Where can I find the K562 cell line DNA sequence?

Hi all, I am trying to use CRISPOR to select target sequences for Crispr/Cas9 knock-in, but an error

1) whats the minimum coverage you would shoot for if you want a good number of hits. My understanding

Hello Meenu, I am facing a similar issue and was wondering whether you have meanwhile proceeded with

Hi, in this case you have no choice but to remove the stop codon of your gene then you can acquire

Hi, everyone! I am in the process of constructing an EGFP knock-in cell line using the CRISPR-HDR

Kindly suggest me any online tool used to design efficient sgRNA negative control Sequences. On Fri,

I'm actually in the process of using it right now. Our lab recently returned to a project that we

I'd say you should use a similar number of non-targeting guides as a control. But interested to

You need to change this line as well, 20 to 18. guide = read_sequence[start_index:(start_index + 20)]

Dear all, we have the last 4 seats available for the course "Introduction to CRISPR for ecology

Hello, fellow researchers, I'm working on a project cloning 11 guide RNAs into a Pxr003 plasmid

Hello, fellow researchers, I'm working on a project cloning 11 guide RNAs into a Pxr003 plasmid

Hi Experts I have been trying to develop the CRISPR/Cas9 lentivirus based whole genome screening for

Hi Gozde from my experience, it's fine to try sanger, However enrichment is necessary, except

Hi Muhammad, I used UNAfold, and it does appear your sticky ends can form a hairpin deltaG = -2.23

Hi,all: We found that many articles used QIAamp DNA blood kit to extract the genome, but we found

Hi Rashu, This depends on what promoter you have used and also where the Gene integrated in the

Hi Julia and the community I have amplified the libraries from almost 30 samples from my screen (

Hello, In our laboratory we have done a positive screening with a drug. The sequencing data with the

Hello, I've noticed a sequence labeled "Terminator" post the second direct repeat in

remove Met on the Cas protein and add the kozak sequence before HA tag. It doesn't change the

Hi Guys, I am having some doubts about the design of the vector and the placement of sgRNA in the

Greetings everyone! I recently modified Cas9 and aim to incorporate it into another plasmid for

Dear All, I am currently exploring strategies to repress transcription via inhibiting RNA polymerase,