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Micro-dissection and integration of long and short reads to create a robust catalog of kidney compartment-specific isoforms
- Publication Year :
- 2021
- Publisher :
- Cold Spring Harbor Laboratory, 2021.
-
Abstract
- Studying isoform expression at the microscopic level has always been a challenging task. A classical example is kidney, where glomerular and tubulo-insterstitial compartments carry out drastically different physiological functions and thus presumably their isoform expression also differs. We aim at developing an experimental and computational pipeline for identifying isoforms at microscopic structure-level. We microdissed glomerular and tubulo-interstitial compartments from healthy human kidney tissues from two cohorts. The two compartments were separately sequenced with the PacBio RS II platform. These transcripts were then validated using transcripts of the same samples by the traditional Illumina RNA-Seq protocol, distinct Illumina RNA-Seq short reads from European Renal cDNA Bank (ERCB) samples, and annotated GENCODE transcript list, thus identifying novel transcripts. We identified 14,739 and 14,259 annotated transcripts, and 17,268 and 13,118 potentially novel transcripts in the glomerular and tubulo-interstitial compartments, respectively. Of note, relying solely on either short or long reads would have resulted in many erroneous identifications. We identified distinct pathways involved in glomerular and tubulointerstitial compartments at the isoform level.We demonstrated the possibility of micro-dissecting a tissue, incorporating both long- and short-read sequencing to identify isoforms for each compartment.
- Subjects :
- Gene isoform
Classical example
Computational biology
Biology
Kidney
urologic and male genital diseases
Cellular and Molecular Neuroscience
Complementary DNA
Genetics
medicine
Humans
Protein Isoforms
Compartment (development)
RNA, Messenger
Molecular Biology
Ecology, Evolution, Behavior and Systematics
Ecology
urogenital system
GENCODE
Gene Expression Profiling
Microscopic level
High-Throughput Nucleotide Sequencing
Human kidney
medicine.anatomical_structure
Computational Theory and Mathematics
Modeling and Simulation
Subjects
Details
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....a7ca653f5f8436376c18f35717fb3460
- Full Text :
- https://doi.org/10.1101/2021.09.07.459298