How can i select best gRNA for CRISPER/Cas9 vector ?

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

How to get the vector of a specific vibration mode in phonon dispersion?

For example, I have this phonon dispersion(Fig. 1), and now I would like get vector of atoms which describes TA1 in P. I know v_sim can show vibrational animation of specific mode, but I do not...

20 February 2021 4,332 4 View

Specify the Burgers vectors for dislocations, which cannot be uniquely identified.?

1. A bcc metal single crystal was sliced along (1 -1 0) plane for TEM study. Assuming that the TEM available is equipped with a goniometer stage which tilts up to 45o. The Burgers vector of...

20 February 2021 2,370 1 View

I am working on image transmission, i converted the image into binary vector, i want to extract the MSB of each pixel ?

i have a vector 1*512 size, i want to take the first value every 8 elements and put these elements at the end of the vector,how to do this on matlab ?

19 February 2021 2,352 5 View

In what ways do fellow reaerchers think that mRNA vaccination may be under CRISPR systems technology application ?

There are many ways for mRNA to intervene with genome expression. This question is raised for experts in CRISPR systems. Any RNA does not stand alone within acell. There are many ways to become...

18 February 2021 9,109 5 View

Does the property of GFP affect a fusion protein's localization?

Hello, I am planning to insert GOI between CaMV 35 promoter and mGFP in pCambia1302 to produce a fusion protein and see subcellular localization of the protein. But the vector says it's membrane...

15 February 2021 4,353 3 View

How much amount of DNA (ligation mix) should I use for transformation?

Hi Fellow Scientists! I am trying to clone my 300 bp insert into a 6 kb vector. I run a few different ligation reactions using (1:3 vector: insert molar ratios) 10, 20,30, and 40 ng/ul vector...

15 February 2021 9,563 6 View

Can I express protein of plant origin in E. coli strain (M15) using pET28a vector systems? Are M15 strains and pET vector systems compatible?

Are E.coli strains M15 and pET28a vector systems compatible? Can I over-express my protein which is of plant origin. Previously I was using BL21 DE3 strains. Was confused to use M15 strains for...

10 February 2021 2,564 3 View

Plasmid amplification in S2 cell?

When co-transfect S2 cell with selection vector (pCoHygro or pCoBlast ) to make a stable cell line, does the gene integrate into the genome or just like E.coli? How do the plasmids amplify within...

09 February 2021 5,423 1 View

Hendrik de Buhr

Everything you will need you will find here: https://www.addgene.org/crispr/

Md Hassanur Rahman

Thank you Hendrik de Buhr

Kailash Lipne

Hey Rahman you can use the CRISPR Direct software to design the gRNA as per your requirement. That will provide you the details regarding the off target effect of sg RNA with different criteria's.

https://crispr.dbcls.jp/

Than you Kailash Lipne

Yuan-Yeu Yau

Documents have suggested that you design a few (ex. 3) sgRNAs for an allele you want to mutate, to increase the chance the DSB is created. First, those sgRNAs can be tested in vitro, then select the best one for using in vivo. This is one way to go.

Isha Monga

You can also use geCRISPR tool (http://bioinfo.imtech.res.in/manojk/gecrispr).. Hope this tool will solve your purpose..

Cheers..!!!

Thank you Yuan-Yeu Yau and Isha Monga.

Olufunke Dosumu

chopchop is also good