Comprehensive profiling of N6-methyladnosine (m6A) readouts reveals novel m6A readers that regulate human embryonic stem cell differentiation [GLORI-seq]
GLORI-Seq is an antibody-independent sequencing, which can detect m6A sites at single-base. The core of GLORI-Seq is to find a catalytic system of glyoxal and nitrite through chemical reaction combination screening, which efficiently deaminate unmethylated adenosine A to form inosine (A-to-I, > 98%). Inosine is read to guanosine (G) in the sequencing process, forming A-to-G conversion. GLORI-Seq enables single base detection of single base m6A by detecting the proportion of “A” in the sequenced read sequence. Overall design: A549, hESCs and JURKAT cells were analyzed with 2 duplications for GLORI-Seq. METTL3-knockdown A549, hESCs and JURKAT cells were analyzed with 2 duplications for GLORI-Seq.