First evidence for TIGR-Tas activity in a photosynthetic eukaryote
- 1Duke Kunshan University
Abstract
TIGR--Tas systems are a recently described class of RNA-guided DNA-targeting systems whose activity has been demonstrated in bacterial and human cells, but not in photosynthetic eukaryotes. Here we tested TIGR--Tas in the green alga Chlamydomonas reinhardtii. Recombinant TaTas and ParTas assembled with in vitro-transcribed tigRNAs cleaved target DNA in a concentration-dependent manner, and electroporated TaTas protein was detected in C. reinhardtii cells. Targeted deep sequencing of the endogenous MAA7 locus recovered indel-containing reads in ParTas-treated cells that were absent from wild-type controls, including a 1-bp deletion located at the position previously reported for ParTas-associated cleavage. Co-delivery of a double-stranded non-homologous oligonucleotide (dsNHO) with a 24-bp duplex and 8-nt 3' overhangs, designed to match the staggered ends generated by TasR-family nucleases, increased the recovery of indel-containing reads for both proteins. The frequency of these events was very low, and the present data do not establish TIGR--Tas as an efficient genome-editing system in this host. They nonetheless provide initial evidence that TIGR--Tas can act on the nuclear genome of a photosynthetic eukaryote, and indicate that dsNHO-assisted recovery of sequence alterations is not restricted to conventional CRISPR--Cas nucleases.
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BibTeX
@misc{jaeyeon2026first,
title = {{First evidence for TIGR-Tas activity in a photosynthetic eukaryote}},
author = {L. Jaeyeon and K. Kyeongjun and R. Hyeonmin and Y. Hyeonsik and D. Kerstin and P. Mid-Eum and Y. Joong-Tak and K. Bon-Kyoung and Y. Ji-Hye and L. Ho-Seok and K. Eun Yu},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.12.750307},
url = {https://www.biorxiv.org/content/10.64898/2026.09.12.750307v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - First evidence for TIGR-Tas activity in a photosynthetic eukaryote
AU - L. Jaeyeon
AU - K. Kyeongjun
AU - R. Hyeonmin
AU - Y. Hyeonsik
AU - D. Kerstin
AU - P. Mid-Eum
AU - Y. Joong-Tak
AU - K. Bon-Kyoung
AU - Y. Ji-Hye
AU - L. Ho-Seok
AU - K. Eun Yu
PY - 2026
DA - 2026/09/12
PB - bioRxiv
DO - 10.64898/2026.09.12.750307
UR - https://www.biorxiv.org/content/10.64898/2026.09.12.750307v1
AB - TIGR--Tas systems are a recently described class of RNA-guided DNA-targeting systems whose activity has been demonstrated in bacterial and human cells, but not in photosynthetic eukaryotes. Here we tested TIGR--Tas in the green alga Chlamydomonas reinhardtii. Recombinant TaTas and ParTas assembled with in vitro-transcribed tigRNAs cleaved target DNA in a concentration-dependent manner, and electroporated TaTas protein was detected in C. reinhardtii cells. Targeted deep sequencing of the endogenous MAA7 locus recovered indel-containing reads in ParTas-treated cells that were absent from wild-type controls, including a 1-bp deletion located at the position previously reported for ParTas-associated cleavage. Co-delivery of a double-stranded non-homologous oligonucleotide (dsNHO) with a 24-bp duplex and 8-nt 3' overhangs, designed to match the staggered ends generated by TasR-family nucleases, increased the recovery of indel-containing reads for both proteins. The frequency of these events was very low, and the present data do not establish TIGR--Tas as an efficient genome-editing system in this host. They nonetheless provide initial evidence that TIGR--Tas can act on the nuclear genome of a photosynthetic eukaryote, and indicate that dsNHO-assisted recovery of sequence alterations is not restricted to conventional CRISPR--Cas nucleases.
ER -