Protocol to obtain targeted transcript sequence data from snake venom samples collected in the Colombian field.

Area of Science:

• Biochemistry
• Molecular Biology
• Pharmacology

Background:

• Snake venoms are complex mixtures of molecules with therapeutic potential.
• Traditional venom studies often require sacrificing animals or using captive specimens, posing ethical and logistical challenges.
• Accessing venom samples in remote or conflict-affected regions, like parts of Colombia, is difficult.

Purpose of the Study:

• To develop and validate a novel protocol for amplifying complete toxin sequences from field-collected snake venom samples.
• To enable the study of venom composition without sacrificing animals, particularly in challenging geographical and political environments.
• To facilitate research on undercharacterized venoms from species like Crotalus durissus cumanensis.

Main Methods:

• Utilized lyophilized Crotalus durissus cumanensis venom as a model system.
• Designed primers targeting conserved regions of Crotalus sp. mRNA and expressed sequence tags (ESTs) to maximize toxin gene amplification.
• Employed molecular techniques for the amplification and sequencing of toxin genes.

Main Results:

• Successfully amplified and sequenced complete toxin genes including Metalloproteinases II, Disintegrins, Disintegrin-Like, Phospholipases A2, C-type Lectins, and Serine proteinases.
• Confirmed the accuracy of amplified sequences by comparison with existing Crotalus sp. databases.
• Demonstrated the efficacy of the protocol using lyophilized venom and field-collected samples.

Conclusions:

• The developed protocol effectively enables the amplification of complete toxin sequences from lyophilized snake venom, including samples collected in the field.
• This strategy overcomes limitations associated with animal sacrifice and difficult sample access, facilitating the study of snake venoms in challenging environments.
• The method supports the characterization of poorly understood venom components and contributes to drug discovery efforts.