RNA editing in flowering plant mitochondria post-transcriptionally alters several hundred nucleotides from C to U, mostly in mRNAs. We recently identified the nuclear encoded gene MEF11 which is involved in RNA editing of three sites in mRNAs coding for subunits of respiratory chain complexes in Arabidopsis thaliana. In the mef11-2 mutant a T-DNA insert alters the C-terminal part of the DYW domain. This insertion blocks RNA editing at sites cox 3-422 and site nad4-124 completely, but only partially at ccb203-344. Transcription analysis shows that RNA editing is not defective due to disturbance of transcripts from the mef11-2 locus. These observations suggest that the enzymatic activity or its recruitment is not compromised by the alteration of the C-terminus of MEF11.
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http://dx.doi.org/10.4161/psb.11095 | DOI Listing |
Kardiol Pol
January 2025
1st Department of Cardiology, Poznan University of Medical Sciences, Poznań, Poland.
PLoS One
December 2024
Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada.
The ability to determine the essentiality of a gene in the protozoan parasite Leishmania is important to identify potential targets for intervention and understanding the parasite biology. CRISPR gene editing technology has significantly improved gene targeting efficiency in Leishmania. There are two commonly used CRISPR gene targeting methods in Leishmania; the stable expression of the gRNA and Cas9 using a plasmid containing a Leishmania ribosomal RNA gene promoter (rRNA-P stable protocol) and the T7 RNA polymerase based transient gRNA expression system in promastigotes stably expressing Cas9 (T7 transient protocol).
View Article and Find Full Text PDFNat Genet
January 2025
Institute of Molecular Oncology, Philipps-University, Marburg, Germany.
The mutational landscape of TP53, a tumor suppressor mutated in about half of all cancers, includes over 2,000 known missense mutations. To fully leverage TP53 mutation status for personalized medicine, a thorough understanding of the functional diversity of these mutations is essential. We conducted a deep mutational scan using saturation genome editing with CRISPR-mediated homology-directed repair to engineer 9,225 TP53 variants in cancer cells.
View Article and Find Full Text PDFNat Commun
January 2025
Interdisciplinary Life Sciences Graduate Programs, University of Texas at Austin, Austin, TX, 78712, USA.
Type II CRISPR endonucleases are widely used programmable genome editing tools. Recently, CRISPR-Cas systems with highly compact nucleases have been discovered, including Cas9d (a type II-D nuclease). Here, we report the cryo-EM structures of a Cas9d nuclease (747 amino acids in length) in multiple functional states, revealing a stepwise process of DNA targeting involving a conformational switch in a REC2 domain insertion.
View Article and Find Full Text PDFSci Data
January 2025
Gakushuin University, Faculty of Science, Department of Life Science, Mejiro 1-5-1, Toshima-ku, Tokyo, 171-8588, Japan.
The wild silk moth, Bombyx mandarina, is the closest relative of the domesticated silk moth, Bombyx mori. National BioResource Project of Japan (NBRP) maintains a B. mandarina strain derived from individuals captured at Sakado (Saitama, Japan) in 1982.
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