CRISPR

about 1 month ago

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...

2 months ago

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

4 months ago

using a protocol from the broad institute, human CRISPR Brunello lentiviral pooled libraries were amplified using STBL4 cells. A gel electrophoresis image shows two adjacent bands near 14 kb (4th and 6th line), and an additional band near 1 kb which is the amp antibiotic resistance region of the plasmid . Attempting double ...

9 months ago

We are trying to subclone a ~4kb PCR fragment into the pCW-puro/blast backbone (~6kb). Both fragments run at expected MW have been gel-purified. The ligation reaction appeared to have worked efficiently, as the backbone only plate had 0-5 colonies, while the insert-containing ligation yield hundreds of colonies. However, wh...

10 months ago

So, let’s talk ethics and gene editing. Designer dogs have been around as long as domestication… We know where that went- ever seen a pug? “Cute” brachycephalic dogs with severe health issues and short lifespans, just for fun. Now, we’ve got CRIPSR because, obviously, we can’t spend generations and generations in-breeding ...

10 months ago

I use count_spacers.py to analyze the NGS results (paired-end). my code is as below: python countspacers.py -no-g -i sgrnainput.csv -f c_1.clean.fq my percentage of guides that matched perfectly is just 43%, and percentage of undetected guides is more than 96%: Number of perfect guide matches: 1223 Number of nonperfect...

10 months ago

I would be highly appreciated if someone could share a detailed protocol (worked) for knocking out genes from any HCC cancer cell lines using px459-v2. Thank you

11 months ago

This piece honestly always puts the work I do in perspective. Especially seeing war and famine go about...it's nice to know that the tech we're using can be truly revolutionary in LITERALLY saving lives. Also...with crop diversification there can be all sorts of interesting new combos.

11 months ago

Saw this cool article on a tomato that was released in Japan (https://academic.oup.com/pcp/article-abstract/63/6/731/6564444?redirectedFrom=fulltext&login=false)... Would love to try this thing :D

12 months ago

I am new to the use of CRISPR libraries and I would like some input regarding a problem we are having. I am. using the GeCKO v2 library according to the Joung et al. Nature protocols paper. After library amplification we amplified the sgRNA targets by PCR using an equimolar mix of the primers foward of the paper and one of ...

dCasRx/13 and splicing

dCasRx/13 and splicing

CRISPR ARRAY - Cas13

CRISPR ARRAY - Cas13

Concerns over additional bands in CRISPR lentiviral libraries amplified using STBL4 cells

Concerns over additional bands in CRISPR lentiviral libraries amplified using STBL4 cells

pCW cloning by RE ligation with weird product

pCW cloning by RE ligation with weird product

Designer Babies

Designer Babies

Low match in my data using count_spacers.py

Low match in my data using count_spacers.py

Gene knockout protocol from HCC cancer cell lines using px459-v2?

Gene knockout protocol from HCC cancer cell lines using px459-v2?

Solving the Food Crisis with CRISPR

Solving the Food Crisis with CRISPR

Launching this CRISPR tomato crop in a japanese market...

Launching this CRISPR tomato crop in a japanese market...

poor quality of reads after library amplification

poor quality of reads after library amplification