Mutant/mutation
The mutant expresses LISP2 C-terminal tagged as follows: Pb-Cre parasites were generated by fusing Cre recombinase with an SV40 nuclear localization signal (NLS) and HA tag on the C-terminus of Lisp2 by CRISPR-mediated insertion of these coding sequences in the endogenous pblisp2 genomic locus.

CRISPR-mediated insertion/tagging was performed using the CRISPR-RGR technology described for mutant RMgm-4633.

Protein (function)
Evidence has been presented that LISP2 plays a role in development of maturing liver schizonts. LISP2 is specifically expressed in (late) liver stages. Evidence has been presented for export of LISP2 into the cytoplasm of the hepatocyte. The protein has been named LISP2 and sequestrin.

Phenotype
Evidence presented that LISP2 tagged with Cre recombinase is exported into hepatocytes and that the Cre-recombinase is active in infected hepatocytes (see below).

Additional information
To demonstrate the extent to which Pb-Cre can ablate target DNA sequences in host hepatocytes, we infected primary hepatocytes derived from Ai14 mice with Pb-Cre. Ai14 mice possess a floxed STOP cassette preventing the transcription of a CAG promoter-driven red fluorescent protein variant, tdTomato. Upon excision of the STOP cassette, tdTomato expression is ‘turned on’ in Ai14 cells. Ai14 hepatocytes infected with Pb-Cre showed robust tdTomato expression, indicating that Pb-Cre can be reliably used to ablate floxed target DNA sequences in Plasmodium-infected cells.

Other mutants