Table 3 Some limitations and solutions in the application of CRISPR/Cas9 gene editing

Off-target mutagenesis

Correct design of sgRNA

D10A-mutated Cas9

SpCas9-HF1 and eSpCas9

[188,189,190]

Identification of the PAM region

SpCas9-NG and xCas9 variants

[191]

P53 activation and apoptosis

Cas9 variants with the ability to create single-strand breaks

[191]

Anti-SpCas9 antibodies

Campylobacter jejuni Cas9 (CjCas9)

[191, 192]

Stimulation of immune responses in delivery by viral vector

Using other delivery method such as nanoparticle

[193]