Joseph Miano

5 Questions 15 Answers 0 Followers

Questions related from Joseph Miano

Is there innate immunity in the mouse zygote?

RNA can elicit innate immunity in cultured cells during CRISPR editing, a fact that further supports the growing use of Cas9-synthetic gRNA as an RNP for superior editing in mouse embryos. But,...

09 September 2018 9,070 0 View

How fast does siRNA work in cells?

A collaborator of mine has shown data where siRNA knockdown OF A TRANSCRIPT occurs as early as 8 hrs post-transfection. I cannot seem to find any similar data online. Has anyone out there...

06 June 2018 9,318 3 View

Does loss of one nucleotide in a loxP site abolish its recognition and cleavage by Cre?

So we made a CRISPR mouse and got both loxP sites on same allele (no small feat). The 5' loxP site is perfect, the 3' loxP site, located 1.5 kb downstream, as lost an "A" nucleotide at the 5' end...

07 July 2015 8,913 5 View

How many conditional alleles in mice have been made with CRISPR?

The original approach from the Jaenisch lab uses two ssoligos and 2 guide RNAs. We were lucky to have obtained one (out of 40 genotyped) mice, though a single base pair deletion outside the CRE...

01 January 1970 9,794 3 View

What conditions and anti-FLAG antibody are proven to work in tissue sections of mouse?

We recently made a FLAG-tagged mouse using 3 component CRISPR. We can do ChIP and Westerns very well. However, detecting the FLAG epitope in mouse sections (either frozen or paraffin) have been...

01 January 1970 8,733 5 View