Research team improves geneediting with chemicalprocess NatureComms
, these RNA epigenetic editing events can be coupled to various cellular signals, which can lead to new synthetic biology strategies in engineering mammalian cells for research or therapeutic purposes.A editors were constructed by replacing dCas13b or METTL3 from pCMV-dCas13-M3nls with ABI or PYL. Typically, PYL or ABI primers ordered from Thermo Fisher Scientific were used to amplify PYL or ABI fragments with Q5 High-Fidelity 2x Master Mix , using PCR instrument .
Finally, the membrane was washed with TBST three times and then imaged on ChemiDocHEK293T cells or Hela cells were seeded in six-well plate and transfected with plasmids at nearly 70% confluence. At 24 h post-transfection, cells were fixed with 4% paraformaldehyde for 10 min. Subsequently, the above-fixed cells were treated with 0.1% Triton™ X-100 for 15 min for permeabilization and then blocked with 1% bovine serum albumin for 1 h at room temperature.
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