Cas13 endonuclease activity depends on the RNA local secondary structure with strong preference for single-stranded (SS) regions. Henceit becomes indispensable to identify the SS regions for effective Cas13 mediated RNA knockdown. We herein present rational gRNA design by integrating experimental ...
. These recombinant proteins were expressed in the HEK293E cell system, which has been used extensively forPlasmodiumprotein expression23, and purified using affinity chromatography. To test for cross-reactivity, we used the antisera to probe Western blots containing a panel of 12 purified recombinant...
Also, Cas9 is a member of a small subset of Cas effectors that need a second trans-acting crRNA (tracrRNA) for gRNA processing and DNA cleavage [11]. Recently, Zhang and his group at MIT and Broad Institute (USA) discovered a new generation of CRISPR nucleases termed as Cpf1 (CRISPR ...
CRISPR and CRISPR-associated (Cas) protein, as components of microbial adaptive immune system, allows biologists to edit genomic DNA in a precise and specific way. CRISPR-Cas systems are classified into two main classes and six types. Cpf1 is a putative type V (class II) CRISPR effector, wh...
Also, Cas9 is a member of a small subset of Cas effectors that need a second trans-acting crRNA (tracrRNA) for gRNA pro- cessing and DNA cleavage [11]. Recently, Zhang and his group at MIT and Broad Institute (USA) discovered a new generation of CRISPR nucleases termed as Cpf1 (...
The heterogeneity of functional cardiomyocytes arises during heart development, which is essential to the complex and highly coordinated cardiac physiological function. Yet the biological and physiological identities and the origin of the specialized cardiomyocyte populations have not been fully comprehended. ...
NOX2 is the prototypical member of the NADPH oxidase NOX superfamily and produces superoxide (O2•−), a key reactive oxygen species (ROS) that is essential in innate and adaptive immunity. Mutations that lead to deficiency in NOX2 activity correlate
Here, we develop spegRNA by introducing same-sense mutations at proper positions in the reverse-transcription template of pegRNA to increase PE’s base-editing efficiency up-to 4,976-fold (on-average 353-fold). We also develop apegRNA by altering the pegRNA secondary structure to increase PE...
Generation of TRC-KO cells by CRISPR/ Cas-9 Nicking Strategy TCR negative versions of Jurkat cell lines were generated using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome engineering. Guide RNAs (gRNA) were designed targeting exon 1 of the TRBC1 gene, choosing the...
(a) The design of the msgRNA-3 construct. The AAVS1.sgRNA scaffold and another irrelevant shRNA were inserted into the msgRNA-2 construct upstream of the VEGF.sgRNA scaffold. (b) The illustration of the RPG surrogate reporter construct based on SSA repair. Similar with the DsRed-eGFP ...