. 2023 Oct 5;31:100733. doi: 10.1016/j.omto.2023.100733

Table 3.

CRISPR screens in other immune cells to enhance their anti-tumor reactivity

Species	Target cells	Loss or gain of function	CRISPR library	Transduction methods	Selection methods	Genes identified	Corresponding proteins	Gene/protein functions	Years and references
Mouse	raw macrophage cell line 264.7	loss of function	self-designed library with 7272 sgRNA targeting classical RBP genes	lentivirus transduction	TNF-α production	Mettl3	METTL3	METTL3-mediated m6A modification of Irakm mRNA accelerates its degradation, resulting in reprogramming macrophages for activation.	Tong et al. (2021)¹¹³
Mouse	cDC1s	loss of function	self-designed functional CRISPR library	retrovirus transduction	OVA cross-presentation	Wdfy4	WDFY4	Loss of function of BEACH domain–containing protein Wdfy4 substantially impaired cross-presentation of cell-associated antigens by cDC1s in mice.	Theisen et al. (2018)¹¹⁴
Human	IL-2 dependent NK cell line NK-92	gain of function	self-designed library with 22 gRNAs using Benchling’s CRISPR Design tool	Cas RNP nucleofection	calcein-AM cytotoxic assay	FCGR3A	FCGR3A	Cas9-mediated promoter insertion effectively reactivated the endogenous FCGR3A and CD226 enhanced NK-92 cytotoxicity.	Huang et al. (2020)¹¹¹
	IL-2 dependent NK cell line NK-92	gain of function		Cas RNP nucleofection	calcein-AM cytotoxic assay	CD226	CD226		Huang et al. (2020)¹¹¹
	moDC	loss of function	Brunello	electroporation of in vitro-assembled Cas9-sgRNA complexes	loss of TNF-α secretion	TLR2	TLR2	TLR2 knockout caused decrease in TNF-α secretion.	Jost et al. (2021)¹¹⁵
	moDC	loss of function	Brunello		loss of TNF-α secretion	PTPN6/SHP-1	PTPN6	PTPN6 knockout strongly increased TNF-α secretion and moderately decreased IL-10 secretion.	Jost et al. (2021)¹¹⁵
	human myeloid cell line U937	loss of function	self-designed 10-sgRNA-per-gene CRISPR-Cas9 deletion library	lentivirus transduction	phagocytosis ability	NHLRC2	NHLRC2	NHLRC2 negatively regulates RHOA, enabling RAC1-mediated cytoskeletal changes that are critical for phagocytosis.	Haney et al. (2018)¹¹⁶
						ELOVL1	ELOVL1	ELOVL1 gene disruption causes significant decrease in phagocytosis.
						TM2D3	TM2D3	TM2D2- and TM2D3-deficient cells show impaired clearance of amyloid-β aggregates.

cDC1, classical dendritic cell; ELOVL1, elongation of very long chain fatty acids protein 1; FCGR3A, Fc gamma receptor IIIa; IL-2, interleukin-2; METTL3, methyltransferase 3; moDC, monocyte-derived dendritic cell; NHLRC2, NHL repeat containing protein 2; PTPN6, protein tyrosine phosphatase non-receptor type 6; TLR2, Toll-like receptor 2; TM2D3, TM2 domain containing 3; WDFY4, Wdfy family member 4.