التفاصيل البيبلوغرافية
| العنوان: |
CRISPR editing methods for constructing atlas loss-of-function mutations. |
| المؤلفون: |
Emily L. Rivard (8708418), Andrew G. Ludwig (11383983), Prajal H. Patel (8708419), Anna Grandchamp (5632529), Sarah E. Arnold (11383986), Alina Berger (11383989), Emilie M. Scott (11383992), Brendan J. Kelly (7329878), Grace C. Mascha (11383995), Erich Bornberg-Bauer (135063), Geoffrey D. Findlay (8708422) |
| سنة النشر: |
2021 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biochemistry, Cell Biology, Genetics, Molecular Biology, Evolutionary Biology, Developmental Biology, Inorganic Chemistry, Infectious Diseases, Environmental Sciences not elsewhere classified, Biological Sciences not elsewhere classified, study provides insight, several independent lineages, proper chromatin condensation, fundamental developmental process, div >< p, cytological analyses showed, complex evolutionary history, based chromatin packaging, 15 million years, identified one gene, gene likely arose, spermatid chromatin condensation, male reproductive tissues, de novo <, drosophila melanogaster <, functional genetic screen, evolved gene required, drosophila <, gene moved, spermatid nuclei |
| الوصف: |
A) Graphical representation of CRISPR loss-of-function strategies. Purple boxes represent the atlas coding region, while gray represents non-coding regions of the gene. Scissors indicate locations where gRNAs targeted Cas9-mediated double-stranded breaks. The deletion allele was generated using two sgRNAs targeting either side of atlas to excise the complete coding region (CDS) and nearly all of the noncoding region. Frameshift alleles were created using one sgRNA targeting a cut at the start of the atlas coding region that was repaired with non-homologous end joining (NHEJ). B) Alignment of mutations generated in frameshift alleles. The gene’s start codon is indicated with dark green shading. Blue shading indicates bases inserted by NHEJ, orange shading indicates NHEJ deletions. All three mutations consist of net insertions or deletions that are non-multiples of three, resulting in premature stop codons indicated with underlining. The mutant alleles retain the possibility of encoding a truncated form of Atlas protein if a downstream start codon (light green shading) is used, since this codon is in-frame with the sequence encoding the protein’s C terminus. C) Predicted protein sequences encoded by the wild-type and frameshift alleles. Blue shading indicates novel amino acids created by NHEJ indel mutations. Gray shading indicates the potential truncated Atlas protein that could be encoded by the frameshift alleles if translation initiated at the downstream start codon indicated in panel B. Such hypothetical, N-terminally truncated forms of Atlas protein would contain amino acids 61–172 of the wild-type protein. Yellow shading indicates the putative nuclear localization signal. (PDF) |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/journal_contribution/CRISPR_editing_methods_for_constructing_i_atlas_i_loss-of-function_mutations_/16568637 |
| DOI: |
10.1371/journal.pgen.1009787.s002 |
| الاتاحة: |
https://doi.org/10.1371/journal.pgen.1009787.s002 |
| Rights: |
CC BY 4.0 |
| رقم الانضمام: |
edsbas.643D4152 |
| قاعدة البيانات: |
BASE |